Helicobacter antigens and corresponding DNA fragments

ABSTRACT

The invention provides Helicobacter polypeptides that can be used in vaccination methods for preventing or treating Helicobacter infection, and polynucleotides that encode these polypeptides.

PRIORITY INFORMATION

[0001] This application is a continuation of, and claims priority from,U.S. Ser. No. 08/749,051, filed Nov. 14, 1996, which is incorporated byreference herein.

FIELD OF THE INVENTION

[0002] The invention relates to Helicobacter antigens and correspondingDNA molecules, which can be used in methods to prevent and treatHelicobacter infection in mammals, such as humans.

BACKGROUND OF THE INVENTION

[0003] Helicobacter is a genus of spiral, gram-negative bacteria thatcolonize the gastrointestinal tracts of mammals. Several speciescolonize the stomach, most notably H. pylori, H. heilmanii, H. felis,and H. mustelae. Although H. pylori is the species most commonlyassociated with human infection, H. heilmanii and H. felis have alsobeen isolated from humans, but at lower frequencies than H. pylori.Helicobacter infects over 50% of adult populations in developedcountries and nearly 100% in developing countries and some Pacific rimcountries, making it one of the most prevalent infections worldwide.

[0004] Helicobacter is routinely recovered from gastric biopsies ofhumans with histological evidence of gastritis and peptic ulceration.Indeed, H. pylori is now recognized as an important pathogen of humans,in that the chronic gastritis it causes is a risk factor for thedevelopment of peptic ulcer diseases and gastric carcinoma. It is thushighly desirable to develop safe and effective vaccines for preventingand treating Helicobacter infection.

[0005] A number of Helicobacter antigens have been characterized orisolated. These include urease, which is composed of two structuralsubunits of approximately 30 and 67 kDa (Hu et al., Infect. Immun.58:992, 1990; Dunn et al., J. Biol. Chem. 265:9464, 1990; Evans et al.,Microbial Pathogenesis 10:15, 1991; Labigne et al., J. Bact., 173:1920,1991); the 87 kDa vacuolar cytotoxin (VacA) (Cover et al., J. Biol.Chem. 267:10570, 1992; Phadnis et al., Infect. Immun. 62:1557, 1994; WO93/18150); a 128 kDa immunodominant antigen associated with thecytotoxin (CagA, also called TagA) (WO 93/18150; U.S. Pat. No.5,403,924); 13 and 58 kDa heat shock proteins HspA and HspB (Suerbaum etal., Mol. Microbiol. 14:959, 1994; WO 93/18150); a 54 kDa catalase(Hazell et al., J. Gen. Microbiol. 137:57, 1991); a 15 kDahistidine-rich protein (Hpn) (Gilbert et al., Infect. Immun. 63:2682,1995); a 20 kDa membrane-associated lipoprotein (Kostrcynska et al., J.Bact. 176:5938, 1994), an 30 kDa outer membrane protein (Bölin et al.,J. Clin. Microbiol. 33:381, 1995); a lactoferrin receptor (FR2,724,936), and several porins, referred to as HopA, HopB, HopC, HopD,and HopE, which have molecular weights of 48-67 kDa (Exner et al.,Infect. Immun. 63:1567, 1995; Doig et al., J. Bact. 177:5447, 1995).

[0006] Some of these proteins have been proposed as potential vaccineantigens. In particular, urease is believed to be a vaccine candidate(WO 94/9823; WO 95/22987; WO 95/3824; Michetti et al., Gastroenterology107:1002, 1994). Nevertheless, it is contemplated that several antigensmay ultimately be necessary in a vaccine.

SUMMARY OF THE INVENTION

[0007] The present invention provides DNA molecules that encodeHelicobacter polypeptides designated HPO101, HPO104, HPO116, HPO121,HPO132, HPO15, HPO18, HPO38, HPO42, HPO45, HPO50, HPO54, HPO57, HPO58,HPO64, HPO70, HPO71, HPO76, HPO7, HPO80, HPO87, HPO95, HPO98, and HPO9,which can be used in methods to prevent, treat, and diagnoseHelicobacter infection. The encoded polypeptides include polypeptideshaving the amino acid sequences shown in SEQ ID NOs:2 to 48 (evennumbers) and polypeptides encoded by DNA inserts found in depositedplasmids (see below, e.g., Example 2). Those skilled in the art willappreciate that the invention also includes DNA molecules that encodemutants and derivatives of such polypeptides, which result from theaddition, deletion, or substitution of non-essential amino acids asdescribed herein. The invention also includes RNA moleculescorresponding to the DNA molecules of the invention.

[0008] In addition to the DNA and RNA molecules, the invention includesthe corresponding polypeptides and monospecific antibodies thatspecifically bind to such polypeptides.

[0009] The present invention has wide application and includesexpression cassettes, vectors, and cells transformed or transfected withthe polynucleotides of the invention. Accordingly, the present inventionprovides (i) a method for producing a polypeptide of the invention in arecombinant host system and related expression cassettes, vectors, andtransformed or transfected cells; (ii) a live vaccine vector, such as apox virus, Salmonella typhimurium, or Vibrio cholerae vector, containinga polynucleotide of the invention, such vaccine vectors being usefulfor, e.g., preventing and treating Helicobacter infection, incombination with a diluent or carrier, and related pharmaceuticalcompositions and associated therapeutic and/or prophylactic methods;(iii) a therapeutic and/or prophylactic method involving administrationof an RNA or DNA molecule of the invention, either in a naked form orformulated with a delivery vehicle, a polypeptide or combination ofpolypeptides, or a monospecific antibody of the invention, and relatedpharmaceutical compositions; (iv) a method for diagnosing the presenceof Helicobacter in a biological sample, which can involve the use of aDNA or RNA molecule, a monospecific antibody, or a polypeptide of theinvention; and (v) a method for purifying a polypeptide of the inventionby antibody-based affinity chromatography.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010]FIG. 1A is a diagrammatic representation of transposon TnMax9,which is a derivative of the TnMax transposon system (Haas et al., Gene130:23-21, 1993). The mini-transposon carries the blaM gene, which isthe β-lactamase gene lacking a promoter and a signal sequence, next tothe inverted repeats (IR) and the M13 forward (M13-FP) and reverse(M13-RP1) primer binding sites. The resolution site (res) and an originof replication (ori_(fd)) are located between the blaM gene and theconstitutive cat_(GC)-resistance gene. The transposase tnpA andresolvase tnpR genes are located outside of the mini-transposon and areunder the control of the inducible P_(trc) promoter. The lacIq geneencodes the Lac repressor.

[0011]FIG. 1B is a diagrammatic representation of plasmid pMin2. pMin2contains a multiple cloning site, the tetracycline resistance gene(tet), an origin of transfer (oriT), an origin of replication(ori_(ColE1)), a transcriptional terminator (t_(fd)), and a weak,constitutive promoter (P_(iga)). H. pylori chromosome fragments wereintroduced into the BglII and ClaI sites of pMin2.

[0012] FIGS. 2A-2E are a series of graphs showing an analysis of some ofthe physical properties of polypeptide HPO76 (SEQ ID NO:36).

[0013] FIGS. 3A-3D are a series of graphs showing an analysis of some ofthe physical properties of polypeptide HPO15 (SEQ ID NO:12).

[0014] FIGS. 4A-4F are a series of graphs showing an analysis of some ofthe physical properties of polypeptide HPO42 (SEQ ID NO:18).

[0015] FIGS. 5A-5D are a series of graphs showing an analysis of some ofthe physical properties of polypeptide HPO50 (SEQ ID NO:22).

[0016] FIGS. 6A-6H are a series of graphs showing an analysis of some ofthe physical properties of polypeptide HPO54 (SEQ ID NO:24).

[0017] FIGS. 7A-7G are a series of graphs showing an analysis of some ofthe physical properties of polypeptide HPO57 (SEQ ID NO:26).

[0018] FIGS. 8A-8G are a series of graphs showing an analysis of some ofthe physical properties of polypeptide HPO64 (SEQ ID NO:30).

DETAILED DESCRIPTION

[0019] In the H. pylori genome, open reading frames (ORFs) encoding fulllength, membrane-associated secreted/excreted polypeptides have beennewly identified. These polypeptides include membrane polypeptidespermanently found in the membrane structure and polypeptides that arepresent in the external vicinity of the membrane. These polypeptides canbe used in vaccination methods for preventing and treating Helicobacterinfection. The ORFs encode secreted polypeptides that can be readilyproduced in their mature form (polypeptides exported through class II orIII secretion pathway) or are initially produced as precursors includinga signal peptide that can be removed in the course ofexcretion/secretion by cleavage at the N-terminal end of the matureform. (The cleavage site is located at the C-terminal end of the signalpeptide, adjacent to the mature form.) In the sequences disclosed in thepresent application, these cleavage sites and accordingly the firstamino acid of the mature polypeptides, were putatively determined.

[0020] According to a first aspect of the invention, there are providedisolated polynucleotides encoding the precursor and mature forms ofHelicobacter HPO101, HPO104, HPO116, HPO121, HPO132, HPO15, HPO18,HPO38, HPO42, HPO45, HPO50, HPO54, HPO57, HPO58, HPO64, HPO70, HPO71,HPO76, HPO7, HPO80, HPO87, HPO95, HPO98, and HPO9.

[0021] An isolated polynucleotide of the invention encodes (i) apolypeptide having an amino acid sequence that is homologous to aHelicobacter amino acid sequence of a polypeptide associated with theHelicobacter membrane, the Helicobacter amino acid sequence beingselected from the group consisting of:

[0022] (a) the amino acid sequences as shown:

[0023] in SEQ ID NO:2, beginning with an amino acid in any one of thepositions from −27 to 5, preferably in position −27 or position 1, andending with an amino acid in position 160 (HPO101);

[0024] in SEQ ID NO:4, beginning with an amino acid in position 1 andending with an amino acid in position 172 (HPO104);

[0025] in SEQ ID NO:6, beginning with an amino acid in any one of thepositions from −17 to 5, preferably in position −17 or position 1, andending with an amino acid in position 169 (HPO116);

[0026] in SEQ ID NO:8, beginning with an amino acid in any one of thepositions from −21 to 5, preferably in position −20 or position 1, andending with an amino acid in position 198 (HPO121);

[0027] in SEQ ID NO:10, beginning with an amino acid in any one of thepositions from −20 to 5, preferably in position −20 or position 1, andending with an amino acid in position 132 (HPO132);

[0028] in SEQ ID NO:12, beginning with an amino acid in 1 to 5,preferably in position 1, and ending with an amino acid in position 114(HPO15);

[0029] in SEQ ID NO:14, beginning with an amino acid in any one of thepositions from −17 to 5, preferably in position −17 or position 1, andending with an amino acid in position 248 (HPO18);

[0030] in SEQ ID NO:16, beginning with an amino acid in any one of thepositions from −40 to 5, preferably in position −40 or position 1, andending with an amino acid in position 74 (HPO38);

[0031] in SEQ ID NO:18, beginning with an amino acid in any one of thepositions from −34 to 5, preferably in position −34 or position 1, andending with an amino acid in position 226 (HPO42);

[0032] in SEQ ID NO:20, beginning with an amino acid in any one of thepositions from −21 to 5, preferably in position −21 or position 1, andending with an amino acid in position 179 (HPO45);

[0033] in SEQ ID NO:22, beginning with an amino acid in any one of thepositions from −33 to 5, preferably in position −33 or position 1, andending with an amino acid in position 114 (HPO50);

[0034] in SEQ ID NO:24, beginning with an amino acid in any one of thepositions from −60 to 5, preferably in position −60 or position 1, andending with an amino acid in position 349 (HPO54);

[0035] in SEQ ID NO:26, beginning with an amino acid in any one of thepositions from −18 to 5, preferably in position −18 or position 1, andending with an amino acid in position 288 (HPO57);

[0036] in SEQ ID NO:28, beginning with an amino acid in any one of thepositions from −21 to 5, preferably in position −21 or position 1, andending with an amino acid in position 150 (HPO58);

[0037] in SEQ ID NO:30, beginning with an amino acid in any one of thepositions from −20 to 5, preferably in position −20 or position 1, andending with an amino acid in position 309 (HPO64);

[0038] in SEQ ID NO:32, beginning with an amino acid in any one of thepositions from −35 to 5, preferably in position −35 or position 1, andending with an amino acid in position 129 (HPO70);

[0039] in SEQ ID NO:34, beginning with an amino acid in any one of thepositions from −19 to 5, preferably in position −19 or position 1, andending with an amino acid in position 153 (HPO71);

[0040] in SEQ ID NO:36, beginning with an amino acid in any one of thepositions from −25 to 5, preferably in position −25 or position 1, andending with an amino acid in position 176 (HPO76);

[0041] in SEQ ID NO:38, beginning with an amino acid in any one of thepositions from −21 to 5, preferably in position −21 or position 1, andending with an amino acid in position 156 (HPO7);

[0042] in SEQ ID NO:40, beginning with an amino acid in position 1 andending with an amino acid in position 144 (HPO80);

[0043] in SEQ ID NO:42, beginning with an amino acid in any one of thepositions from −20 to 5, preferably in position −20 or position 1, andending with an amino acid in position 152 (HPO87);

[0044] in SEQ ID NO:44, beginning with an amino acid in any one of thepositions from −31 to 5, preferably in position −31 or position 1, andending with an amino acid in position 112 (HPO95);

[0045] in SEQ ID NO:46, beginning with an amino acid in any one of thepositions from −20 to 5, preferably in position −20 or position 1, andending with an amino acid in position 91 (HPO98);

[0046] in SEQ ID NO:48, beginning with an amino acid in any one of thepositions from −21 to 5, preferably in position −21 or position 1, andending with an amino acid in position 129 (HPO9); and

[0047] (b) the precursor or mature amino acid sequences encoded by theH. pylori DNA inserts found in American Type Culture Collection depositnumbers HPO76 (98197), HPO18 (98210), HPO121 (98201), HPO45 (98208),HPO101 (98198), HPO116 (98200), HPO7 (98211), HPO104 (98199), HPO15(98214), HPO58 (98206), HPO132 (98202), HPO9 (98203), HPO38 (98204),HPO87 (98205), HPO71 (98217), HPO70 (98219), HPO80 (98215), HPO95(98216), HPO98 (98218), HPO57 (98220), HPO50 (98207), HPO64 (98213),HPO54 (98212), and HPO42 (98209); or (ii) a derivative of thepolypeptide.

[0048] The term “isolated polynucleotide” is defined as a polynucleotideremoved from the environment in which it naturally occurs. For example,a naturally-occurring DNA molecule present in the genome of a livingbacteria or as part of a gene bank is not isolated, but the samemolecule separated from the remaining part of the bacterial genome, as aresult of, e.g., a cloning event (amplification), is isolated.Typically, an isolated DNA molecule is free from DNA regions (e.g.,coding regions) with which it is immediately contiguous at the 5′ or 3′end, in the naturally occurring genome. Such isolated polynucleotidescould be part of a vector or a composition and still be isolated in thatsuch a vector or composition is not part of its natural environment.

[0049] A polynucleotide of the invention can be in the form of RNA orDNA (e.g., cDNA, genomic DNA, or synthetic DNA), or modifications orcombinations thereof. The DNA can be double-stranded or single-stranded,and, if single-stranded, can be the coding strand or the non-coding(anti-sense) strand. The sequence that encodes a polypeptide of theinvention as shown in SEQ ID NOs:2 to 48 (even numbers), or encoded by adeposited DNA molecule, can be (a) the coding sequence as shown in SEQID NOs:1 to 47 (odd numbers), (b) the coding sequence of a deposited DNAmolecule of the invention (see below); (c) a ribonucleotide sequencederived by transcription of (a) or (b); or (d) a different codingsequence; this latter, as a result of the redundancy or degeneracy ofthe genetic code, encodes the same polypeptides as the DNA molecules ofwhich the nucleotide sequences are illustrated in SEQ ID NOs:1 to 47(odd numbers) or the deposited DNA molecules of the invention.

[0050] Advantageously, the polypeptide is naturally secreted or excretedby Helicobacter felis, H. mustelae, H. heilmanii, or H. pylori; thelatter being preferred.

[0051] By “polypeptide” or “protein” is meant any chain of amino acids,regardless of length or post-translational modification (e.g.,glycosylation or phosphorylation). Both terms are used interchangeablyin the present application.

[0052] By “homologous amino acid sequence” is meant an amino acidsequence that differs from an amino acid sequence shown in SEQ IDNOs:2-48 (even numbers) or encoded by a deposited DNA molecule of theinvention, only by one or more conservative amino acid substitutions, orby one or more non-conservative amino acid substitutions, deletions, oradditions located at positions at which they do not destroy the specificantigenicity of the polypeptide.

[0053] Preferably, such a sequence is at least 75%, more preferably 80%,and most preferably 90% identical to an amino acid sequence shown in SEQID NOs:2 to 48 (even numbers) or encoded by a deposited DNA molecule ofthe invention.

[0054] Homologous amino acid sequences include sequences that areidentical or substantially identical to an amino acid sequence as shownin SEQ ID NOs:2 to 48 (even numbers) or encoded by a deposited DNAmolecule of the invention. By “amino acid sequence substantiallyidentical” is meant a sequence that is at least 90%, preferably 95%,more preferably 97%, and most preferably 99% identical to an amino acidsequence of reference and that preferably differs from the sequence ofreference, if at all, by a majority of conservative amino acidsubstitutions.

[0055] Conservative amino acid substitutions typically includesubstitutions among amino acids of the same class. These classesinclude, for example, amino acids having uncharged polar side chains,such as asparagine, glutamine, serine, threonine, and tyrosine; aminoacids having basic side chains, such as lysine, arginine, and histidine;amino acids having acidic side chains, such as aspartic acid andglutamic acid; and amino acids having nonpolar side chains, such asglycine, alanine, valine, leucine, isoleucine, proline, phenylalanine,methionine, tryptophan, and cysteine.

[0056] As an illustration of substitutive variations, particularexamples are provided as follows. In the sequence shown in SEQ ID NO:4,the lysine in position 96 can be substituted with asparagine, glutamine,isoleucine, threonine, glutamic acid, or arginine; the asparagines inpositions 120 and 123 can be substituted with isoleucine, threonine,lysine, serine, tyrosine, or asparagine; the lysines in positions 125,128, and 144 can be substituted with asparagine, glutamine, isoleucine,threonine, glutamic acid, or arginine; or the proline in position 150can be substituted with serine, threonine, alanine, leucine, arginine,or histidine. In the sequence shown in SEQ ID NO:8, the leucine inposition 115 can be substituted with phenylalanine, isoleucine, valine,proline, histidine, or arginine. In the sequence shown in SEQ ID NO:10,the arginine in position 107 can be substituted with glycine, theasparagine in position 118 can be substituted with isoleucine,threonine, or serine; or the proline in position 130 can be substitutedwith serine, threonine, alanine, leucine, arginine, or histidine. In thesequence shown in SEQ ID NO:12, the asparagine in position 17 can besubstituted with isoleucine, threonine, or serine. In the sequence shownin SEQ ID NO:12, the asparagine in position 17 can be , substituted withisoleucine, threonine, or serine. In the sequence shown in SEQ ID NO:40,the asparagine in position 33 can be substituted with isoleucine,threonine, or serine, and the phenylalanine in position 128 can besubstituted with serine, tyrosine, or cysteine. In the sequence shown inSEQ ID NO:50, the glutamine in position 10 can be substituted withleucine, proline, or arginine; the leucine in position 26 can besubstituted with phenylalanine, and the arginine in position 127 can besubstituted with glycine.

[0057] Homology is typically measured using sequence analysis software(e.g., Sequence Analysis Software Package of the Genetics ComputerGroup, University of Wisconsin Biotechnology Center, 1710 UniversityAvenue, Madison, Wis. 53705). Similar amino acid sequences are alignedto obtain the maximum degree of homology (i.e., identity). To this end,it may be necessary to artificially introduce gaps into the sequence.Once the optimal alignment has been set up, the degree of homology(i.e., identity) is established by recording all of the positions inwhich the amino acids of both sequences are identical, relative to thetotal number of positions.

[0058] Homologous polynucleotide sequences are defined in a similar way.Preferably, a homologous sequence is one that is at least 45%, morepreferably 60%, and most preferably 85% identical to (i) a codingsequence of SEQ ID NOs:1 to 47 (odd numbers), or (ii) a coding sequenceof a deposited DNA molecule of the invention.

[0059] Polypeptides having a sequence homologous to one of the sequencesshown in SEQ ID NOs:2 to 48 (even numbers), include naturally-occurringallelic variants, as well as mutants or any other non-naturallyoccurring variants that are analogous in terms of antigenicity, to apolypeptide having a sequence as shown in SEQ ID NOs:2 to 48 (evennumbers) or encoded by a deposited DNA molecule of the invention.

[0060] As is known in the art, an allelic variant is an alternate formof a polypeptide that is characterized as having a substitution,deletion, or addition of one or more amino acids that does not alter thebiological function of the polypeptide. By “biologic function” is meantthe function of the polypeptide in the cells in which it naturallyoccurs, even if the function is not necessary for the growth or survivalof the cells. For example, the biological function of a porin is toallow the entry into cells of compounds present in the extracellularmedium. The biological function is distinct from the antigenic function.A polypeptide can have more than one biological function.

[0061] Allelic variants are very common in nature. For example, abacterial species, e.g., H. pylori, is usually represented by a varietyof strains that differ from each other by minor allelic variations.Indeed, a polypeptide that fulfills the same biological function indifferent strains can have an amino acid sequence that is not identicalin each of the strains. Such an allelic variation may be equallyreflected at the polynucleotide level.

[0062] Support for the use of allelic variants of polypeptide antigenscomes from, e.g., studies of the Helicobacter urease antigen. The aminoacid sequence of Helicobacter urease varies widely from species tospecies, yet cross-species protection occurs, indicating that the ureasemolecule, when used as an immunogen, is highly tolerant of amino acidvariations. Even among different strains of the single species H.pylori, there are amino acid sequence variations.

[0063] For example, although the amino acid sequences of the UreA andUreB subunits of H. pylori and H. felis ureases differ from one anotherby 26.5% and 11.8%, respectively (Ferrero et al., Molecular Microbiology9(2):323-333, 1993), it has been shown that H. pylori urease protectsmice from H. felis infection (Michetti et al., Gastroenterology107:1002-1011, 1994). In addition, it has been shown that the individualstructural subunits of urease, UreA and UreB, which contain distinctamino acid sequences, are both protective antigens against Helicobacterinfection (Michetti et al., supra). Similarly, Cuenca et al.(Gastroenterology 110: 1770-1775, 1996) showed that therapeuticimmunization of H. mustelae-infected ferrets with H. pylori urease waseffective at eradicating H. mustelae infection. Further, several ureasevariants have been reported to be effective vaccine antigens, including,e.g., recombinant UreA+UreB apoenzyme expressed from pORV142 (UreA andUreB sequences derived from H. pylori strain CPM630; Lee et al., J.Infect. Dis. 172:161-172, 1995); recombinant UreA+UreB apoenzymeexpressed from pORV214 (UreA and UreB sequences differ from H. pyloristrain CPM630 by one and two amino acid changes, respectively; Lee etal., supra, 1995); a UreA-glutathione-S-transferase fusion protein (UreAsequence from H. pylori strain ATCC 43504; Thomas et al., ActaGastro-Enterologica Belgica, 56:54, September 1993); UreA+UreBholoenzyme purified from H. pylori strain NCTC11637 (Marchetti et al.,Science 267:1655-1658, 1995); a UreA-MBP fusion protein (UreA from H.pylori strain 85P; Ferrero et al., Infection and Immunity 62:4981-4989,1994); a UreB-MBP fusion protein (UreB from H. pylori strain 85P;Ferrero et al., supra); a UreA-MBP fusion protein (UreA from H. felisstrain ATCC 49179; Ferrero et al., supra); a UreB-MBP fusion protein(UreB from H. felis strain ATCC 49179; Ferrero et al., supra); and a 37kD fragment of UreB containing amino acids 220-569 (Dore-Davin et al.,“A 37 kD fragment of UreB is sufficient to confer protection againstHelicobacter felis infection in mice”). Finally, Thomas et al. (supra)showed that oral immunization of mice with crude sonicates of H. pyloriprotected mice from subsequent challenge with H. felis.

[0064] Polynucleotides, e.g., DNA molecules, encoding allelic variantscan easily be retrieved by polymerase chain reaction (PCR) amplificationof genomic bacterial DNA extracted by conventional methods. Thisinvolves the use of synthetic oligonucleotide primers matching upstreamand downstream of the 5′ and 3′ ends of the encoding domain. Suitableprimers can be designed according to the nucleotide sequence informationprovided in SEQ ID NOs:1 to 47 (odd numbers). Typically, a primer canconsist of 10 to 40, preferably 15 to 25 nucleotides. It may be alsoadvantageous to select primers containing C and G nucleotides in aproportion sufficient to ensure efficient hybridization; e.g., an amountof C and G nucleotides of at least 40%, preferably 50% of the totalnucleotide amount.

[0065] As an example, primers useful for cloning by PCR a DNA moleculeencoding a polypeptide having the amino acid sequence of HPO76 (SEQ IDNO:36), or encoded by the corresponding deposited DNA molecule(pMin2/76; HPO76, ATCC Deposit Number 98197), are shown in SEQ ID NO:83(matching at the 5′ end) and in SEQ ID NO:84 (matching at the 3′ end).Experimental conditions for carrying out PCR can readily be determinedby one skilled in the art and an illustration of carrying out PCR isprovided in Example 1.

[0066] Thus, the first aspect of the invention includes (i) isolated DNAmolecules that can be amplified and/or cloned by polymerase chainreaction from a Helicobacter, e.g., H. pylori, genome, using either:

[0067] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:49, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:50;

[0068] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:51, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:52;

[0069] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:53, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:54;

[0070] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:55, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:56;

[0071] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:57, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:58;

[0072] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:59, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:60;

[0073] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:61, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:62;

[0074] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:63, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:64;

[0075] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:65, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:66;

[0076] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:67, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:68;

[0077] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:69, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:70;

[0078] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:71, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:72;

[0079] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:73, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:74;

[0080] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:75, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:76;

[0081] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:77, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:78;

[0082] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:79, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:80;

[0083] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:81, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:82;

[0084] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:83, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:84;

[0085] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:85, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:86;

[0086] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:87, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:88;

[0087] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:89, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:90;

[0088] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:91, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:93;

[0089] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:95, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:94;

[0090] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:97, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:96; or

[0091] A 5′ oligonucleotide primer having a sequence as shown in SEQ IDNO:99, and a 3′ oligonucleotide primer having a sequence in SEQ IDNO:98; and

[0092] (ii) isolated DNA molecules encoding the mature forms of thepolypeptides encoded by the DNA molecules amplified as above.

[0093] In the sequences provided in SEQ ID NOs:49 to 96, the letter “N”denotes a restriction site that contains, typically, 4 to 6 nucleotides.Restriction sites can be selected by those skilled in the art so thatthe amplified DNA can be conveniently cloned into an appropriatelydigested plasmid.

[0094] Useful homologs that do not naturally occur can be designed usingknown methods for identifying regions of an antigen that are likely tobe tolerant of amino acid sequence changes and/or deletions. Forexample, sequences of the antigen from different species can be comparedto identify conserved sequences.

[0095] Polypeptide derivatives that are encoded by polynucleotides ofthe invention include, e.g., fragments, polypeptides having largeinternal deletions derived from full-length polypeptides, and fusionproteins.

[0096] Polypeptide fragments of the invention can be derived from apolypeptide having a sequence homologous to any of the sequences shownin SEQ ID NOs:2 to 48 (even numbers) or encoded by a deposited DNAmolecule of the invention (see below, e.g., Example 2), to the extentthat the fragments retain the substantial antigenicity of the parentpolypeptide (specific antigenicity). Polypeptide derivatives can also beconstructed by large internal deletions that remove a substantial partof the parent polypeptide, while retaining specific antigenicity.Generally, polypeptide derivatives should be about at least 12 aminoacids in length to maintain antigenicity. Advantageously, they can be atleast 20 amino acids, preferably at least 50 amino acids, morepreferably at least 75 amino acids, and most preferably at least 100amino acids in length.

[0097] Useful polypeptide derivatives, e.g., polypeptide fragments, canbe designed using computer-assisted analysis of amino acid sequences inorder to identify sites in protein antigens having potential assurface-exposed, antigenic regions (Hughes et al., Infect. Immun.60(9):3497, 1992).

[0098] Computer-assisted analysis of some polypeptides of the inventionis illustrated in FIGS. 2 to 8, which are graphs showing some of thephysical properties of polypeptides HPO76 (SEQ ID NO:36), HPO15 (SEQ IDNO:12), HPO42 (SEQ ID NO:18), HPO50 (SEQ ID NO:22), HPO54 (SEQ IDNO:24), HPO57 (SEQ ID NO:26), and HPO64 (SEQ ID NO:30). The graphs wereprepared using the Laser Gene Program from DNA Star, and include, e.g.,hydrophilicity, antigenic index, and intensity index plots. Alsoincluded in the graphs are spots showing homologies with known proteinmotifs, such as the T-cell recognition motif and the majorhistocompatibility complex (MHC) IA and IE regions of mice. One skilledin the art can readily use the information provided in such plots toselect peptide fragments for use as vaccine antigens. For example,fragments spanning regions of the plots in which the antigenic index isrelatively high can be selected. One can also select fragments spanningregions in which both the antigenic index and the intensity plots arerelatively high. Fragments containing conserved sequences, particularlyhydrophilic conserved sequences, can also be selected.

[0099] Polypeptide fragments and polypeptides having large internaldeletions can be used for revealing epitopes that are otherwise maskedin the parent polypeptide and that may be of importance for inducing aprotective T cell-dependent immune response. Deletions can also removeimmunodominant regions of high variability among strains.

[0100] It is an accepted practice in the field of immunology to usefragments and variants of protein immunogens as vaccines, as all that isrequired to induce an immune response to a protein is a small (e.g., 8to 10 amino acid) immunogenic region of the protein. This has been donefor a number of vaccines against pathogens other than Helicobacter. Forexample, short synthetic peptides corresponding to surface-exposedantigens of pathogens such as murine mammary tumor virus (peptidecontaining 11 amino acids; Dion et al., Virology 179:474-477, 1990),Semliki Forest virus (peptide containing 16 amino acids; Snijders etal., J. Gen. Virol. 72:557-565, 1991), and canine parvovirus (2overlapping peptides, each containing 15 amino acids; Langeveld et al.,Vaccine 12(15): 1473-1480, 1994) have been shown to be effective vaccineantigens against their respective pathogens.

[0101] Polynucleotides encoding polypeptide fragments and polypeptideshaving large internal deletions can be constructed using standardmethods (see, e.g., Ausubel et al., Current Protocols in MolecularBiology, John Wiley & Sons Inc., 1994), for example, by PCR, includinginverse PCR, by restriction enzyme treatment of the cloned DNAmolecules, or by the method of Kunkel et al. (Proc. Natl. Acad. Sci.U.S.A. 82:448, 1985; biological material available at Stratagene).

[0102] A polypeptide derivative can also be produced as a fusionpolypeptide that contains a polypeptide or a polypeptide derivative ofthe invention fused, e.g., at the N- or C-terminal end, to any otherpolypeptide (hereinafter referred to as a peptide tail). Such a productcan be easily obtained by translation of a genetic fusion, i.e., ahybrid gene. Vectors for expressing fusion polypeptides are commerciallyavailable, such as the pMal-c2 or pMal-p2 systems of New EnglandBiolabs, in which the peptide tail is a maltose binding protein, theglutathione-S-transferase system of Pharmacia, or the His-Tag systemavailable from Novagen. These and other expression systems provideconvenient means for further purification of polypeptides andderivatives of the invention.

[0103] Another particular example of fusion polypeptides included ininvention includes a polypeptide or polypeptide derivative of theinvention fused to a polypeptide having adjuvant activity, such as,e.g., subunit B of either cholera toxin or E. coli heat-labile toxin.Several possibilities are can be used for achieving fusion. First, thepolypeptide of the invention can be fused to the N-, or preferably, tothe C-terminal end of the polypeptide having adjuvant activity. Second,a polypeptide fragment of the invention can be fused within the aminoacid sequence of the polypeptide having adjuvant activity.

[0104] As stated above, the polynucleotides of the invention encodeHelicobacter polypeptides in precursor or mature form. They can alsoencode hybrid precursors containing heterologous signal peptides, whichcan mature into polypeptides of the invention. By “heterologous signalpeptide” is meant a signal peptide that is not found in thenaturally-occurring precursor of a polypeptide of the invention.

[0105] A polynucleotide of the invention, having a homologous codingsequence, hybridizes, preferably under stringent conditions, to apolynucleotide having a sequence as shown in SEQ ID NOs:1 to 47 (oddnumbers) or to an insert of a deposited DNA molecule (see below, e.g.,Example 2). Hybridization procedures are, e.g., described in Ausubel etal., supra; Silhavy et al. (Experiments with Gene Fusions, Cold SpringHarbor Laboratory Press, 1984); Davis et al. (A Manual for GeneticEngineering: Advanced Bacterial Genetics, Cold Spring Harbor LaboratoryPress, 1980). Important parameters that can be considered for optimizinghybridization conditions are reflected in a formula that allowscalculation of a critical value, the melting temperature above which twocomplementary DNA strands separate from each other (Casey et al., Nucl.Acid Res. 4:1539, 1997). This formula is as follows: Tm=81.5+0.5×(%G+C)+1.6 log (positive ion concentration) −0.6×(% formamide). Underappropriate stringency conditions, hybridization temperature (Th) isapproximately 20 to 40° C., 20 to 25° C., or, preferably 30 to 40° C.below the calculated Tm. Those skilled in the art will understand thatoptimal temperature and salt conditions can be readily determinedempirically in preliminary experiments using conventional procedures.

[0106] For example, stringent conditions can be achieved, both forpre-hybridizing and hybridizing incubations, (i) within 4-16 hours at42° C., in 6× SSC containing 50% formamide or (ii) within 4-16 hours at65° C. in an aqueous 6× SSC solution (1 M NaCl, 0.1 M sodium citrate (pH7.0)).

[0107] For polynucleotides containing 30 to 600 nucleotides, the aboveformula is used and then is corrected by subtracting (600/polynucleotidesize in base pairs). Stringency conditions are defined by a Th that is 5to 10° C. below Tm.

[0108] Hybridization conditions with oligonucleotides shorter than 20-30bases do not exactly follow the rules set forth above. In such cases,the formula for calculating the Tm is as follows: Tm=4×(G+C)+2(A+T). Forexample, an 18 nucleotide fragment of 50% G+C would have an approximateTm of 54° C.

[0109] Plasmids containing nucleic acids encoding HPO101, HPO104,HPO116, HPO121, HPO132, HPO15, HPO18, HPO38, HPO42, HPO45, HPO50, HPO54,HPO57, HPO58, HPO64, HPO70, HPO71, HPO76, HPO7, HPO80, HPO87, HPO95,HPO98, and HPO9 were deposited in E. coli strain DH5α under the BudapestTreaty, with the American Type Culture Collection (ATCC; Rockville, Md.)on Oct. 9, 1996 and were designated with accession numbers listed belowin Example 2. These plasmids were derived from pMin2 by insertion of agenomic DNA BglII-ClaI fragment from H. pylori strain P1 or P12 into thevector. Each of the inserts is disrupted by the presence of transposonTnMax9 (Kahrs et al., Gene 167:53, 1995). The locations of insertion ofthe transposon in each of the deposited clones (see below) are betweenthe nucleotides indicated in parentheses after the name of each clone,as follows: HPO101 (497-498), HPO104 (428-429), HPO116 (433-444), HPO121(463-464), HPO132 (408-409), HPO18 (226-227), HPO38 (347-348), HPO42(372-373), HPO45 (299-300), HPO50 (29-293), HPO54 (351-352), HPO57(266-267), HPO58 (434-435), HPO64 (224-225), HPO70 (114-115), HPO71(274-275), HPO76 (412-413), HPO7 (349-350), HPO80 (105-106), HPO87(26-27), HPO95 (64-65), HPO98 (43-44), and HPO9 (346-347). As isdiscussed further below in Example 2, DNA molecules lacking thetransposon can be amplified from the plasmids using standard PCRtechniques, including inverse and recombinant PCR (see, e.g., PCRprotocols: A Guide to Methods and Applications (1990) Innis et al.,Eds., Academic Press), so that the full-length H. pylori insert isreconstituted.

[0110] A polynucleotide molecule of the invention, containing RNA, DNA,or modifications or combinations thereof, can have various applications.For example, a DNA molecule can be used (i) in a process for producingthe encoded polypeptide in a recombinant host system, (ii) in theconstruction of vaccine vectors such as pox viruses, which are furtherused in methods and compositions for preventing and/or treatingHelicobacter infection, (iii) as a vaccine agent (as well as an RNAmolecule), in a naked form or formulated with a delivery vehicle and,(iv) in the construction of attenuated Helicobacter strains that canover-express a polynucleotide of the invention or express it in anon-toxic, mutated form.

[0111] According to a second aspect of the invention, there is thereforeprovided (i) an expression cassette containing a DNA molecule of theinvention placed under the control of the elements required forexpression, in particular under the control of an appropriate promoter;(ii) an expression vector containing an expression cassette of theinvention; (iii) a procaryotic or eucaryotic cell transformed ortransfected with an expression cassette and/or vector of the invention,as well as (iv) a process for producing a polypeptide or polypeptidederivative encoded by a polynucleotide of the invention, which involvesculturing a procaryotic or eucaryotic cell transformed or transfectedwith an expression cassette and/or vector of the invention, underconditions that allow expression of the DNA molecule of the inventionand, recovering the encoded polypeptide or polypeptide derivative fromthe cell culture.

[0112] A recombinant expression system can be selected from procaryoticand eucaryotic hosts. Eucaryotic hosts include yeast cells (e.g.,Saccharomyces cerevisiae or Pichia Pastoris), mammalian cells (e.g.,COS1, NIH3T3, or JEG3 cells), arthropods cells (e.g., Spodopterafrugiperda (SF9) cells), and plant cells. Preferably, a procaryotic hostsuch as E. coli is used. Bacterial and eucaryotic cells are availablefrom a number of different sources to those skilled in the art, e.g.,the American Type Culture Collection (ATCC; Rockville, Md.).

[0113] The choice of the expression system depends on the featuresdesired for the expressed polypeptide. For example, it may be useful toproduce a polypeptide of the invention in a particular lipidated form orany other form.

[0114] The choice of the expression cassette will depend on the hostsystem selected as well as the features desired for the expressedpolypeptide. Typically, an expression cassette includes a promoter thatis functional in the selected host system and can be constitutive orinducible; a ribosome binding site; a start codon (ATG) if necessary, aregion encoding a signal peptide, e.g., a lipidation signal peptide; aDNA molecule of the invention; a stop codon; and optionally a 3′terminal region (translation and/or transcription terminator). Thesignal peptide-encoding region is adjacent to the polynucleotide of theinvention and placed in proper reading frame. The signalpeptide-encoding region can be homologous or heterologous to the DNAmolecule encoding the mature polypeptide and can be specific to thesecretion apparatus of the host used for expression. The open readingframe constituted by the DNA molecule of the invention, solely ortogether with the signal peptide, is placed under the control of thepromoter so that transcription and translation occur in the host system.Promoters, signal peptide encoding regions are widely known andavailable to those skilled in the art and includes, for example, thepromoter of Salmonella typhimurium (and derivatives) that is inducibleby arabinose (promoter araB) and is functional in Gram-negative bacteriasuch as E. coli (as described in U.S. Pat. No. 5,028,530, and in Cagnonet al., Protein Engineering 4(7):843, 1991); the promoter of the gene ofbacteriophage T7 encoding RNA polymerase, that is functional in a numberof E. coli strains expressing T7 polymerase (described in U.S. Pat. No.4,952,496); OspA lipidation signal peptide; and RlpB lipidation signalpeptide (Takase et al., J. Bact. 169:5692, 1987).

[0115] The expression cassette is typically part of an expressionvector, which is selected for its ability to replicate in the chosenexpression system. Expression vectors (e.g., plasmids or viral vectors)can be chosen from those described in Pouwels et al. (Cloning Vectors: ALaboratory Manual 1985, Supp. 1987). They can be purchased from variouscommercial sources.

[0116] Methods for transforming/transfecting host cells with expressionvectors will depend on the host system selected as described in Ausubelet al., supra.

[0117] Upon expression, a recombinant polypeptide of the invention (or apolypeptide derivative) is produced and remains in the intracellularcompartment, is secreted/excreted in the extracellular medium or in theperiplasmic space, or is embedded in the cellular membrane. Thepolypeptide can then be recovered in a substantially purified form fromthe cell extract or from the supernatant after centrifugation of therecombinant cell culture. Typically, the recombinant polypeptide can bepurified by antibody-based affinity purification or by any other methodthat can be readily adapted by a person skilled in the art, such as bygenetic fusion to a small affinity binding domain. Antibody-basedaffinity purification methods are also available for purifying apolypeptide of the invention extracted from a Helicobacter strain.Antibodies useful for purifying by immunoaffinity the polypeptides ofthe invention can be obtained as described below.

[0118] A polynucleotide of the invention can also be useful in thevaccine field, e.g., for achieving DNA vaccination. There are two majorpossibilities, either using a viral or bacterial host as gene deliveryvehicle (live vaccine vector) or administering the gene in a free form,e.g., inserted into a plasmid. Therapeutic or prophylactic efficacy of apolynucleotide of the invention can be evaluated as described below.

[0119] Accordingly, in a third aspect of the invention, there isprovided (i) a vaccine vector such as a pox virus, containing a DNAmolecule of the invention, placed under the control of elements requiredfor expression; (ii) a composition of matter containing a vaccine vectorof the invention, together with a diluent or carrier; particularly,(iii) a pharmaceutical composition containing a therapeutically orprophylactically effective amount of a vaccine vector of the invention;(iv) a method for inducing an immune response against Helicobacter in amammal (e.g., a human; alternatively, the method can be used inveterinary applications for treating or preventing Helicobacterinfection of animals, e.g., cats or birds), which involves administeringto the mammal an immunogenically effective amount of a vaccine vector ofthe invention to elicit an immune response, e.g., a protective ortherapeutic immune response to Helicobacter; and particularly, (v) amethod for preventing and/or treating a Helicobacter (e.g., H. pylori,H. felis, H. mustelae, or H. heilmanii) infection, which involvesadministering a prophylactic or therapeutic amount of a vaccine vectorof the invention to an individual in need. Additionally, the thirdaspect of the invention encompasses the use of a vaccine vector of theinvention in the preparation of a medicament for preventing and/ortreating Helicobacter infection.

[0120] A vaccine vector of the invention can express one or severalpolypeptides or derivatives of the invention, as well as at least oneadditional Helicobacter antigen such as a urease apoenzyme or a subunit,fragment, homolog, mutant, or derivative thereof. In addition, it canexpress a cytokine, such as interleukin-2 (IL-2) or interleukin-12(IL-12), which enhances the immune response (adjuvant effect). Thus, avaccine vector can include an additional DNA molecule encoding, e.g.,urease subunit A, B, or both, or a cytokine, placed under the control ofelements required for expression in a mammalian cell.

[0121] Alternatively, a composition of the invention can include severalvaccine vectors, each of them being capable of expressing a polypeptideor derivative of the invention. A composition can also contain a vaccinevector capable of expressing an additional Helicobacter antigen such asurease apoenzyme, a subunit, fragment, homolog, mutant, or derivativethereof; or a cytokine such as IL-2 or IL-12.

[0122] In vaccination methods for treating or preventing infection in amammal, a vaccine vector of the invention can be administered by anyconventional route in use in the vaccine field, particularly, to amucosal (e.g., ocular, intranasal, oral, gastric, pulmonary, intestinal,rectal, vaginal, or urinary tract) surface or via the parenteral (e.g.,subcutaneous, intradermal, intramuscular, intravenous, orintraperitoneal) route. Preferred routes depend upon the choice of thevaccine vector. The administration can be achieved in a single dose orrepeated at intervals. The appropriate dosage depends on variousparameters understood by skilled artisans such as the vaccine vectoritself, the route of administration or the condition of the mammal to bevaccinated (weight, age and the like).

[0123] Live vaccine vectors available in the art include viral vectorssuch as adenoviruses and pox viruses as well as bacterial vectors, e.g.,Shigella, Salmonella, Vibrio cholerae, Lactobacillus, Bacille bilié deCalmette-Guérin (BCG), and Streptococcus.

[0124] An example of an adenovirus vector, as well as a method forconstructing an adenovirus vector capable of expressing a DNA moleculeof the invention, are described in U.S. Pat. No. 4,920,209. Pox virusvectors that can be used include, e.g., vaccinia and canary pox virus,described in U.S. Pat. Nos. 4,722,848 and 5,364,773, respectively (alsosee, e.g., Tartaglia et al., Virology 188:217, 1992) for a descriptionof a vaccinia virus vector; and Taylor et al, Vaccine 13:539, 1995, fora reference of a canary pox). Pox virus vectors capable of expressing apolynucleotide of the invention can be obtained by homologousrecombination as described in Kieny et al., Nature 312:163, 1984, sothat the polynucleotide of the invention is inserted in the viral genomeunder appropriate conditions for expression in mammalian cells.Generally, the dose of vaccine viral vector, for therapeutic orprophylactic use, can be of from about 1×10⁴ to about 1×10¹¹,advantageously from about 1×10⁷ to about 1×10¹⁰, preferably of fromabout 1×10⁷ to about 1×10⁹ plaque-forming units per kilogram.Preferably, viral vectors are administered parenterally; for example, in3 doses, 4 weeks apart. Those skilled in the art recognize that it ispreferable to avoid adding a chemical adjuvant to a compositioncontaining a viral vector of the invention and thereby minimizing theimmune response to the viral vector itself.

[0125] Non-toxicogenic Vibrio cholerae mutant strains that are useful asa live oral vaccine are described in Mekalanos et al., Nature 306:551,1983, and U.S. Pat. No. 4,882,278 (strain in which a substantial amountof the coding sequence of each of the two ctxA alleles has been deletedso that no functional cholerae toxin is produced); WO 92/11354 (strainin which the irgA locus is inactivated by mutation; this mutation can becombined in a single strain with ctxA mutations); and WO 94/1533(deletion mutant lacking functional ctxA and attRS1 DNA sequences).These strains can be genetically engineered to express heterologousantigens, as described in WO 94/19482. An effective vaccine dose of aVibrio cholerae strain capable of expressing a polypeptide orpolypeptide derivative encoded by a DNA molecule of the invention cancontain, e.g., about 1×10⁵ to about 1×10⁹, preferably about 1×10⁶ toabout 1×10⁸ viable bacteria in an appropriate volume for the selectedroute of administration. Preferred routes of administration include allmucosal routes; most preferably, these vectors are administeredintranasally or orally.

[0126] Attenuated Salmonella typhimurium strains, genetically engineeredfor recombinant expression of heterologous antigens or not, and theiruse as oral vaccines are described in Nakayama et al. (Bio/Technology6:693, 1988) and WO 92/11361. Preferred routes of administration includeall mucosal routes; most preferably, these vectors are administeredintranasally or orally.

[0127] Others bacterial strains useful as vaccine vectors are describedin High et al., EMBO 11:1991, 1992, and Sizemore et al., Science270:299, 1995 (Shigella flexneri); Medaglini et al., Proc. Natl. Acad.Sci. U.S.A. 92:6868, 1995 (Streptococcus gordonii); and Flynn, Cell.Mol. Biol. 40 (suppl. I):31, 1994, WO 88/6626, WO 90/0594, WO 91/13157,WO 92/1796, and WO 92/21376 (Bacille Calmette Guerin).

[0128] In bacterial vectors, polynucleotide of the invention can beinserted into the bacterial genome or can remain in a free state,carried on a plasmid.

[0129] An adjuvant can also be added to a composition containing avaccine bacterial vector. A number of adjuvants are known to thoseskilled in the art. Preferred adjuvants can be selected from the listprovided below.

[0130] According to a fourth aspect of the invention, there is alsoprovided (i) a composition of matter containing a polynucleotide of theinvention, together with a diluent or carrier; (ii) a pharmaceuticalcomposition containing a therapeutically or prophylactically effectiveamount of a polynucleotide of the invention; (iii) a method for inducingan immune response against Helicobacter, in a mammal, by administeringto the mammal, an immunogenically effective amount of a polynucleotideof the invention to elicit an immune response, e.g., a protective immuneresponse to Helicobacter; and particularly, (iv) a method for preventingand/or treating a Helicobacter (e.g., H. pylori, H. felis, H. mustelae,or H. heilmanii) infection, by administering a prophylactic ortherapeutic amount of a polynucleotide of the invention to an individualin need. Additionally, the fourth aspect of the invention encompassesthe use of a polynucleotide of the invention in the preparation of amedicament for preventing and/or treating Helicobacter infection. Thefourth aspect of the invention preferably includes the use of a DNAmolecule placed under conditions for expression in a mammalian cell,e.g., in a plasmid that is unable to replicate in mammalian cells and tosubstantially integrate in a mammalian genome.

[0131] Polynucleotides (DNA or RNA) of the invention can also beadministered as such to a mammal for vaccine, e.g., therapeutic orprophylactic, purpose. When a DNA molecule of the invention is used, itcan be in the form of a plasmid that is unable to replicate in amammalian cell and unable to integrate in the mammalian genome.Typically, a DNA molecule is placed under the control of a promotersuitable for expression in a mammalian cell. The promoter can functionubiquitously or tissue-specifically. Examples of non-tissue specificpromoters include the early Cytomegalovirus (CMV) promoter (described inU.S. Pat. No. 4,168,062) and the Rous Sarcoma Virus promoter (describedin Norton et al., Molec. Cell Biol. 5:281, 1985). The desmin promoter(Li et al., Gene 78:243, 1989, Li et al., J. Biol. Chem. 266:6562, 1991,and Li et al., J. Biol. Chem. 268:10403, 1993) is tissue-specific anddrives expression in muscle cells. More generally, useful vectors aredescribed, i.a., WO 94/21797 and Hartikka et al., Human Gene Therapy7:1205, 1996.

[0132] For DNA/RNA vaccination, the polynucleotide of the invention canencode a precursor or a mature form. When it encodes a precursor form,the precursor form can be homologous or heterologous. In the lattercase, a eucaryotic leader sequence can be used, such as the leadersequence of the tissue-type plasminogen factor (tPA).

[0133] A composition of the invention can contain one or severalpolynucleotides of the invention. It can also contain at least oneadditional polynucleotide encoding another Helicobacter antigen such asurease subunit A, B, or both; or a fragment, derivative, mutant, oranalog thereof. A polynucleotide encoding a cytokine, such asinterleukin-2 (IL-2) or interleukin-12 (IL-12), can also be added to thecomposition so that the immune response is enhanced. These additionalpolynucleotides are placed under appropriate control for expression.Advantageously, DNA molecules of the invention and/or additional DNAmolecules to be included in the same composition, can be carried in thesame plasmid.

[0134] Standard techniques of molecular biology for preparing andpurifying polynucleotides can be used in the preparation ofpolynucleotide therapeutics of the invention. For use as a vaccine, apolynucleotide of the invention can be formulated according to variousmethods.

[0135] First, a polynucleotide can be used in a naked form, free of anydelivery vehicles, such as anionic liposomes, cationic lipids,microparticles, e.g., gold microparticles, precipitating agents, e.g.,calcium phosphate, or any other transfection-facilitating agent. In thiscase, the polynucleotide can be simply diluted in a physiologicallyacceptable solution, such as sterile saline or sterile buffered saline,with or without a carrier. When present, the carrier preferably isisotonic, hypotonic, or weakly hypertonic, and has a relatively lowionic strength, such as provided by a sucrose solution, e.g., a solutioncontaining 20% sucrose.

[0136] Alternatively, a polynucleotide can be associated with agentsthat assist in cellular uptake. It can be, i.a., (i) complemented with achemical agent that modifies the cellular permeability, such asbupivacaine (see, e.g., WO 94/16737), (ii) encapsulated into liposomes,or (iii) associated with cationic lipids or silica, gold, or tungstenmicroparticles.

[0137] Anionic and neutral liposomes are well known in the art (see,e.g., Liposomes: A Practical Approach, RPC New Ed, IRL press (1990), fora detailed description of methods for making liposomes) and are usefulfor delivering a large range of products, including polynucleotides.

[0138] Cationic lipids are also known in the art and are commonly usedfor gene delivery. Such lipids include Lipofectin™ also known as DOTMA(N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride), DOTAP(1,2-bis(oleyloxy)-3-(trimethylammonio)propane), DDAB(dimethyldioctadecylammonium bromide), DOGS (dioctadecylamidologlycylspermine) and cholesterol derivatives such as DC-Chol (3beta-(N-(N′,N′-dimethyl aminomethane)-carbamoyl) cholesterol). Adescription of these cationic lipids can be found in EP 187,702, WO90/11092, U.S. Pat. No. 5,283,185, WO 91/15501, WO 95/26356, and U.S.Pat. No. 5,527,928. Cationic lipids for gene delivery are preferablyused in association with a neutral lipid such as DOPE (dioleylphosphatidylethanolamine), as, for example, described in WO 90/11092.

[0139] Other transfection-facilitating compounds can be added to aformulation containing cationic liposomes. A number of them aredescribed in, e.g., WO 93/18759, WO 93/19768, WO 94/25608, and WO95/2397. They include, i.a., spermine derivatives useful forfacilitating the transport of DNA through the nuclear membrane (see, forexample, WO 93/18759) and membrane-permeabilizing compounds such asGALA, Gramicidine S, and cationic bile salts (see, for example, WO93/19768).

[0140] Gold or tungsten microparticles can also be used for genedelivery, as described in WO 91/359, WO 93/17706, and Tang et al.(Nature 356:152, 1992). In this case, the microparticle-coatedpolynucleotides can be injected via intradermal or intraepidermal routesusing a needleless injection device (“gene gun”), such as thosedescribed in U.S. Pat. Nos. 4,945,050, 5,015,580, and WO 94/24263.

[0141] The amount of DNA to be used in a vaccine recipient depends,e.g., on the strength of the promoter used in the DNA construct, theimmunogenicity of the expressed gene product, the condition of themammal intended for administration (e.g., the weight, age, and generalhealth of the mammal), the mode of administration, and the type offormulation. In general, a therapeutically or prophylactically effectivedose from about 1 μg to about 1 mg, preferably, from about 10 μg toabout 800 μg and, more preferably, from about 25 μg to about 250 μg, canbe administered to human adults. The administration can be achieved in asingle dose or repeated at intervals.

[0142] The route of administration can be any conventional route used inthe vaccine field. As general guidance, a polynucleotide of theinvention can be administered via a mucosal surface, e.g., an ocular,intranasal, pulmonary, oral, intestinal, rectal, vaginal, and urinarytract surface; or via a parenteral route, e.g., by an intravenous,subcutaneous, intraperitoneal, intradermal, intraepidermal, orintramuscular route. The choice of the administration route will dependon, e.g., the formulation that is selected. A polynucleotide formulatedin association with bupivacaine is advantageously administered intomuscles. When a neutral or anionic liposome or a cationic lipid, such asDOTMA or DC-Chol, is used, the formulation can be advantageouslyinjected via intravenous, intranasal (aerosolization), intramuscular,intradermal, and subcutaneous routes. A polynucleotide in a naked formcan advantageously be administered via the intramuscular, intradermal,or sub-cutaneous routes.

[0143] Although not absolutely required, such a composition can alsocontain an adjuvant. If so, a systemic adjuvant that does not requireconcomitant administration in order to exhibit an adjuvant effect ispreferable such as, e.g., QS21, which is described in U.S. Pat. No.5,057,546.

[0144] The sequence information provided in the present applicationenables the design of specific nucleotide probes and primers that can beuseful in diagnosis. Accordingly, in a fifth aspect of the invention,there is provided a nucleotide probe or primer having a sequence foundin or derived by degeneracy of the genetic code from a sequence shown inSEQ ID NO:1 to 47 (odd numbers).

[0145] The term “probe” as used in the present application refers to DNA(preferably single stranded) or RNA molecules (or modifications orcombinations thereof) that hybridize under the stringent conditions, asdefined above, to nucleic acid molecules having sequences homologous tothose shown in SEQ ID NOs:1 to 47 (odd numbers), or to a complementaryor anti-sense sequence. Generally, probes are significantly shorter thanfull-length sequences shown in SEQ ID NOs:1 to 47 (odd numbers); forexample, they can contain from about 5 to about 100, preferably fromabout 10 to about 80 nucleotides. In particular, probes have sequencesthat are at least 75%, preferably at least 85%, more preferably 95%homologous to a portion of a sequence as shown in SEQ ID NOs:1 to 47(odd numbers) or that are complementary to such sequences. Probes cancontain modified bases such as inosine, methyl-5-deoxycytidine,deoxyuridine, dimethylamino-5-deoxyuridine, or diamino-2,6-purine. Sugaror phosphate residues can also be modified or substituted. For example,a deoxyribose residue can be replaced by a polyamide (Nielsen et al.,Science 254:1497, 1991) and phosphate residues can be replaced by estergroups such as diphosphate, alkyl, arylphosphonate and phosphorothioateesters. In addition, the 2′-hydroxyl group on ribonucleotides can bemodified by including, e.g., alkyl groups.

[0146] Probes of the invention can be used in diagnostic tests, ascapture or detection probes. Such capture probes can be conventionallyimmobilized on a solid support, directly or indirectly, by covalentmeans or by passive adsorption. A detection probe can be labeled by adetection marker selected from radioactive isotopes; enzymes such asperoxidase, alkaline phosphatase, and enzymes able to hydrolyze achromogenic, fluorogenic, or luminescent substrate; compounds that arechromogenic, fluorogenic, or luminescent; nucleotide base analogs; andbiotin.

[0147] Probes of the invention can be used in any conventionalhybridization technique, such as dot blot (Maniatis et al., MolecularCloning: A Laboratory Manual (1982) Cold Spring Harbor Laboratory Press,Cold Spring Harbor, N.Y.), Southern blot (Southern, J. Mol. Biol.98:503, 1975), northern blot (identical to Southern blot to theexception that RNA is used as a target), or the sandwich technique (Dunnet al., Cell 12:23, 1977). The latter technique involves the use of aspecific capture probe and/or a specific detection probe with nucleotidesequences that at least partially differ from each other.

[0148] A primer is usually a probe of about 10 to about 40 nucleotidesthat is used to initiate enzymatic polymerization of DNA in anamplification process (e.g., PCR), in an elongation process, or in areverse transcription method. In a diagnostic method involving PCR,primers can be labeled.

[0149] Thus, the invention also encompasses (i) a reagent containing aprobe of the invention for detecting and/or identifying the presence ofHelicobacter in a biological material; (ii) a method for detectingand/or identifying the presence of Helicobacter in a biologicalmaterial, in which (a) a sample is recovered or derived from thebiological material, (b) DNA or RNA is extracted from the material anddenatured, and (c) exposed to a probe of the invention, for example, acapture, detection probe or both, under stringent hybridizationconditions, such that hybridization is detected; and (iii) a method fordetecting and/or identifying the presence of Helicobacter in abiological material, in which (a) a sample is recovered or derived fromthe biological material, (b) DNA is extracted therefrom, (c) theextracted DNA is primed with at least one, and preferably two, primersof the invention and amplified by polymerase chain reaction, and (d) theamplified DNA fragment is produced.

[0150] As previously mentioned, polypeptides that can be produced uponexpression of the newly identified open reading frames are usefulvaccine agents.

[0151] Therefore, a sixth aspect of the invention features asubstantially purified polypeptide or polypeptide derivative having anamino acid sequence encoded by a polynucleotide of the invention.

[0152] A “substantially purified polypeptide” is defined as apolypeptide that is separated from the environment in which it naturallyoccurs and/or that is free of the majority of the polypeptides that arepresent in the environment in which it was synthesized. For example, asubstantially purified polypeptide is free from cytoplasmicpolypeptides. A substantiall purified polypeptide can be, for example,at least 60%, 70%, 80%, 90%, 95%, or 100% pure, with respect to, forexample, other Helicobacter components. Those skilled in the art willunderstand that the polypeptides of the invention can be purified from anatural source, i.e., a Helicobacter strain, or can be produced byrecombinant means.

[0153] Homologous polypeptides or polypeptide derivatives encoded bypolynucleotides of the invention can be screened for specificantigenicity by testing cross-reactivity with an antiserum raisedagainst the polypeptide of reference having an amino acid sequence asshown in SEQ ID NOs:2 to 48 (even numbers) or encoded by one of thedeposited DNA molecules. Briefly, a monospecific hyperimmune antiserumcan be raised against a purified reference polypeptide as such or as afusion polypeptide, for example, an expression product of MBP, GST, orHis-tag systems or a synthetic peptide predicted to be antigenic. Thehomologous polypeptide or derivative screened for specific antigenicitycan be produced as such or as a fusion polypeptide. In this latter caseand if the antiserum is also raised against a fusion polypeptide, twodifferent fusion systems are employed. Specific antigenicity can bedetermined according to a number of methods, including Western blot(Towbin et al., Proc. Natl. Acad. Sci. U.S.A. 76:4350, 1979), dot blot,and ELISA, as described below.

[0154] In a Western blot assay, the product to be screened, either as apurified preparation or a total E. coli extract, is submitted toSDS-Page electrophoresis as described by Laemmli (Nature 227:680, 1970).After transfer to a nitrocellulose membrane, the material is furtherincubated with the monospecific hyperimmune antiserum diluted in therange of dilutions from about 1:50 to about 1:5000, preferably fromabout 1:100 to about 1:500. Specific antigenicity is shown once a bandcorresponding to the product exhibits reactivity at any of the dilutionsin the above range.

[0155] In an ELISA assay, the product to be screened is preferably usedas the coating antigen. A purified preparation is preferred, although awhole cell extract can also be used. Briefly, about 100 μl of apreparation at about 10 μg protein/ml are distributed into wells of a96-well polycarbonate ELISA plate. The plate is incubated for 2 hours at37° C. then overnight at 4° C. The plate is washed with phosphate buffersaline (PBS) containing 0.05% Tween 20 (PBS/Tween buffer). The wells aresaturated with 250 μl PBS containing 1% bovine serum albumin (BSA) toprevent non-specific antibody binding. After 1 hour of incubation at 37°C., the plate is washed with PBS/Tween buffer. The antiserum is seriallydiluted in PBS/Tween buffer containing 0.5% BSA. 100 μl of dilutions areadded per well. The plate is incubated for 90 minutes at 37° C., washedand evaluated according to standard procedures. For example, a goatanti-rabbit peroxidase conjugate is added to the wells when specificantibodies were raised in rabbits. Incubation is carried out for 90minutes at 37° C. and the plate is washed. The reaction is developedwith the appropriate substrate and the reaction is measured bycolorimetry (absorbance measured spectrophotometrically). Under theabove experimental conditions, a positive reaction is shown once an O.D.value of 1.0 is associated with a dilution of at least about 1:50,preferably of at least about 1:500.

[0156] In a dot blot assay, a purified product is preferred, although awhole cell extract can also be used. Briefly, a solution of the productat about 100 μg/ml is serially two-fold diluted in 50 mM Tris-HCl (pH7.5). 100 μl of each dilution are applied to a nitrocellulose membrane0.45 μm set in a 96-well dot blot apparatus (Biorad). The buffer isremoved by applying vacuum to the system. Wells are washed by additionof 50 mM Tris-HCl (pH 7.5) and the membrane is air-dried. The membraneis saturated in blocking buffer (50 mM Tris-HCl (pH 7.5) 0.15 M NaCl, 10μg/L skim milk) and incubated with an antiserum dilution from about 1:50to about 1:5000, preferably about 1:500. The reaction is revealedaccording to standard procedures. For example, a goat anti-rabbitperoxidase conjugate is added to the wells when rabbit antibodies areused. Incubation is carried out 90 minutes at 37° C. and the blot iswashed. The reaction is developed with the appropriate substrate andstopped. The reaction is measured visually by the appearance of acolored spot, e.g., by colorimetry. Under the above experimentalconditions, a positive reaction is shown once a colored spot isassociated with a dilution of at least about 1:50, preferably of atleast about 1:500.

[0157] Therapeutic or prophylactic efficacy of a polypeptide orderivative of the invention can be evaluated as described below.

[0158] According to a seventh aspect of the invention, there is provided(i) a composition of matter containing a polypeptide of the inventiontogether with a diluent or carrier; in particular, (ii) a pharmaceuticalcomposition containing a therapeutically or prophylactically effectiveamount of a polypeptide of the invention; (iii) a method for inducing animmune response against Helicobacter in a mammal, by administering tothe mammal an immunogenically effective amount of a polypeptide of theinvention to elicit an immune response, e.g., a protective immuneresponse to Helicobacter; and particularly, (iv) a method for preventingand/or treating a Helicobacter (e.g., H. pylori, H. felis, H. mustelae,or H. heilmanii) infection, by administering a prophylactic ortherapeutic amount of a polypeptide of the invention to an individual inneed. Additionally, the seventh aspect of the invention encompasses theuse of a polypeptide of the invention in the preparation of a medicamentfor preventing and/or treating Helicobacter infection.

[0159] The immunogenic compositions of the invention can be administeredby any conventional route in use in the vaccine field, in particular toa mucosal (e.g., ocular, intranasal, pulmonary, oral, gastric,intestinal, rectal, vaginal, or urinary tract) surface or via theparenteral (e.g., subcutaneous, intradermal, intramuscular, intravenous,or intraperitoneal) route. The choice of the administration routedepends upon a number of parameters, such as the adjuvant associatedwith the polypeptide. For example, if a mucosal adjuvant is used, theintranasal or oral route will be preferred and if a lipid formulation oran aluminum compound is used, the parenteral route will be preferred. Inthe latter case, the subcutaneous or intramuscular route is mostpreferred. The choice can also depend upon the nature of the vaccineagent. For example, a polypeptide of the invention fused to CTB or LTBwill be best administered to a mucosal surface.

[0160] A composition of the invention can contain one or severalpolypeptides or derivatives of the invention. It can also contain atleast one additional Helicobacter antigen such as the urease apoenzymeor a subunit, fragment, homolog, mutant, or derivative thereof.

[0161] For use in a composition of the invention, a polypeptide orderivative thereof can be formulated into or with liposomes, preferablyneutral or anionic liposomes, microspheres, ISCOMS, orvirus-like-particles (VLPs) to facilitate delivery and/or enhance theimmune response. These compounds are readily available to one skilled inthe art; for example, see Liposomes: A Practical Approach (supra).

[0162] Adjuvants other than liposomes and the like can also be used andare known in the art. An appropriate selection can conventionally bemade by those skilled in the art, for example, from the list providedbelow.

[0163] Administration can be achieved in a single dose or repeated asnecessary at intervals as can be determined by one skilled in the art.For example, a priming dose can be followed by three booster doses atweekly or monthly intervals. An appropriate dose depends on variousparameters including the recipient (e.g., adult or infant), theparticular vaccine antigen, the route and frequency of administration,the presence/absence or type of adjuvant, and the desired effect (e.g.,protection and/or treatment), as can be determined by one skilled in theart. In general, a vaccine antigen of the invention can be administeredby a mucosal route in an amount from about 10 μg to about 500 mg,preferably from about 1 mg to about 200 mg. For the parenteral route ofadministration, the dose usually should not exceed about 1 mg,preferably about 100 μg.

[0164] When used as vaccine agents, polynucleotides and polypeptides ofthe invention can be used sequentially as part of a multistepimmunization process. For example, a mammal can be initially primed witha vaccine vector of the invention such as a pox virus, e.g., via theparenteral route, and then boosted twice with the polypeptide encoded bythe vaccine vector, e.g., via the mucosal route. In another example,liposomes associated with a polypeptide or derivative of the inventioncan also be used for priming, with boosting being carried out mucosallyusing a soluble polypeptide or derivative of the invention incombination with a mucosal adjuvant (e.g., LT).

[0165] A polypeptide derivative of the invention is also useful as adiagnostic reagent for detecting the presence of anti-Helicobacterantibodies, e.g., in a blood sample. Such polypeptides are about 5 toabout 80, preferably about 10 to about 50 amino acids in length and canbe labeled or unlabeled, depending upon the diagnostic method.Diagnostic methods involving such a reagent are described below.

[0166] Upon expression of a DNA molecule of the invention, a polypeptideor polypeptide derivative is produced and can be purified using knownlaboratory techniques. For example, the polypeptide or polypeptidederivative can be produced as a fusion protein containing a fused tailthat facilitates purification. The fusion product can be used toimmunize a small mammal, e.g., a mouse or a rabbit, in order to raiseantibodies against the polypeptide or polypeptide derivative(monospecific antibodies). The eighth aspect of the invention thusprovides a monospecific antibody that binds to a polypeptide orpolypeptide derivative of the invention.

[0167] By “monospecific antibody” is meant an antibody that is capableof reacting with a unique naturally-occuring Helicobacter polypeptide.An antibody of the invention can be polyclonal or monoclonal.Monospecific antibodies can be recombinant, e.g., chimeric (e.g.,constituted by a variable region of murine origin associated with ahuman constant region), humanized (a human immunoglobulin constantbackbone together with hypervariable region of animal, e.g., murine,origin), and/or single chain. Both polyclonal and monospecificantibodies can also be in the form of immunoglobulin fragments, e.g.,F(ab)′2 or Fab fragments. The antibodies of the invention can be of anyisotype, e.g., IgG or IgA, and polyclonal antibodies can be of a singleisotype or can contain a mixture of isotypes.

[0168] The antibodies of the invention, which are raised to apolypeptide or polypeptide derivative of the invention, can be producedand identified using standard immunological assays, e.g., Western blotanalysis, dot blot assay, or ELISA (see, e.g., Coligan et al., CurrentProtocols in Immunology (1994) John Wiley & Sons, Inc., New York, N.Y.).The antibodies can be used in diagnostic methods to detect the presenceof a Helicobacter antigen in a sample, such as a biological sample. Theantibodies can also be used in affinity chromatography methods forpurifying a polypeptide or polypeptide derivative of the invention. Asis discussed further below, such antibodies can be used in prophylacticand therapeutic passive immunization methods.

[0169] Accordingly, a ninth aspect of the invention provides (i) areagent for detecting the presence of Helicobacter in a biologicalsample that contains an antibody, polypeptide, or polypeptide derivativeof the invention; and (ii) a diagnostic method for detecting thepresence of Helicobacter in a biological sample, by contacting thebiological sample with an antibody, a polypeptide, or a polypeptidederivative of the invention, such that an immune complex is formed, andby detecting such complex to indicate the presence of Helicobacter inthe sample or the organism from which the sample is derived.

[0170] Those skilled in the art will understand that the immune complexis formed between a component of the sample and the antibody,polypeptide, or polypeptide derivative, whichever is used, and that anyunbound material can be removed prior to detecting the complex. As canbe easily understood, a polypeptide reagent is useful for detecting thepresence of anti-Helicobacter antibodies in a sample, e.g., a bloodsample, while an antibody of the invention can be used for screening asample, such as a gastric extract or biopsy, for the presence ofHelicobacter polypeptides.

[0171] For use in diagnostic applications, the reagent (i.e., theantibody, polypeptide, or polypeptide derivative of the invention) canbe in a free state or immobilized on a solid support, such as a tube, abead, or any other conventional support used in the field.Immobilization can be achieved using direct or indirect means. Directmeans include passive adsorption (non-covalent binding) or covalentbinding between the support and the reagent. By “indirect means” ismeant that an anti-reagent compound that interacts with a reagent isfirst attached to the solid support. For example, if a polypeptidereagent is used, an antibody that binds to it can serve as ananti-reagent, provided that it binds to an epitope that is not involvedin the recognition of antibodies in biological samples. Indirect meanscan also employ a ligand-receptor system, for example, a molecule suchas a vitamin can be grafted onto the polypeptide reagent and thecorresponding receptor can be immobilized on the solid phase. This isillustrated by the biotin-streptavidin system. Alternatively, indirectmeans can be used, e.g., by adding to the reagent a peptide tail,chemically or by genetic engineering, and immobilizing the grafted orfused product by passive adsorption or covalent linkage of the peptidetail.

[0172] According to a tenth aspect of the invention, there is provided aprocess for purifying, from a biological sample, a polypeptide orpolypeptide derivative of the invention, which involves carrying outantibody-based affinity chromatography with the biological sample,wherein the antibody is a monospecific antibody of the invention.

[0173] For use in a purification process of the invention, the antibodycan be polyclonal or monospecific, and preferably is of the IgG type.Purified IgGs can be prepared from an antiserum using standard methods(see, e.g., Coligan et al., supra). Conventional chromatographysupports, as well as standard methods for grafting antibodies, aredisclosed in, e.g., Antibodies: A Laboratory Manual, D. Lane, E. Harlow,Eds. (1988).

[0174] Briefly, a biological sample, such as an H. pylori extract,preferably in a buffer solution, is applied to a chromatographymaterial, preferably equilibrated with the buffer used to dilute thebiological sample so that the polypeptide or polypeptide derivative ofthe invention (i.e., the antigen) is allowed to adsorb onto thematerial. The chromatography material, such as a gel or a resin coupledto an antibody of the invention, can be in batch form or in a column.The unbound components are washed off and the antigen is then elutedwith an appropriate elution buffer, such as a glycine buffer or a buffercontaining a chaotropic agent, e.g., guanidine HCl, or high saltconcentration (e.g., 3 M MgCl₂). Eluted fractions are recovered and thepresence of the antigen is detected, e.g., by measuring the absorbanceat 280 nm.

[0175] An antibody of the invention can be screened for therapeuticefficacy as described as follows. According to an eleventh aspect of theinvention, there is provided (i) a composition of matter containing amonospecific antibody of the invention, together with a diluent orcarrier; (ii) a pharmaceutical composition containing a therapeuticallyor prophylactically effective amount of a monospecific antibody of theinvention, and (iii) a method for treating or preventing a Helicobacter(e.g., H. pylori, H. felis, H. mustelae, or H. heilmanii) infection, byadministering a therapeutic or prophylactic amount of a monospecificantibody of the invention to an individual in need. Additionally, theeleventh aspect of the invention encompasses the use of a monospecificantibody of the invention in the preparation of a medicament fortreating or preventing Helicobacter infection.

[0176] To this end, the monospecific antibody can be polyclonal ormonoclonal, preferably of the IgA isotype (predominantly). In passiveimmunization, the antibody can be administered to a mucosal surface of amammal, e.g., the gastric mucosa, e.g., orally or intragastrically,advantageously, in the presence of a bicarbonate buffer. Alternatively,systemic administration, not requiring a bicarbonate buffer, can becarried out. A monospecific antibody of the invention can beadministered as a single active component or as a mixture with at leastone monospecific antibody specific for a different Helicobacterpolypeptide. The amount of antibody and the particular regimen used canreadily be determined by those skilled in the art. For example, dailyadministration of about 100 to 1,000 mg of antibodies over one week, orthree doses per day of about 100 to 1,000 mg of antibodies over two orthree days, can be an effective regimens for most purposes.

[0177] Therapeutic or prophylactic efficacy can be evaluated usingstandard methods in the art, e.g., by measuring induction of a mucosalimmune response or induction of protective and/or therapeutic immunity,using, e.g., the H. felis mouse model and the procedures described inLee et al. (Eur. J. Gastroenterology and Hepatology 7:303, 1995) or Leeet al. (J. Infect. Dis. 172:161, 1995). Those skilled in the art willrecognize that the H. felis strain of the model can be replaced withanother Helicobacter strain. For example, the efficacy of DNA moleculesand polypeptides from H. pylori is preferably evaluated in a mouse modelusing an H. pylori strain. Protection can be determined by comparing thedegree of Helicobacter infection in the gastric tissue (assessed byurease activity, bacterial counts or gastritis) to that of a controlgroup. Protection is shown when infection is reduced by comparison tothe control group. Such an evaluation can be made for polynucleotides,vaccine vectors, polypeptides and derivatives thereof, as well asantibodies of the invention.

[0178] For example, various doses of an antibody of the invention can beadministered to the gastric mucosa of mice previously challenged with anH. pylori strain, as described, e.g., in Lee et al (supra). Then, afteran appropriate period of time, the bacterial load of the mucosa isestimated by assessing the urease activity, as compared to a control.Reduced urease activity indicates that the antibody is therapeuticallyeffective.

[0179] Adjuvants useful in any of the vaccine compositions describedabove are as follows.

[0180] Adjuvants for parenteral administration include aluminumcompounds, such as aluminum hydroxide, aluminum phosphate, and aluminumhydroxy phosphate. The antigen can be precipitated with, or adsorbedonto, the aluminum compound according to standard protocols. Otheradjuvants, such as RIBI (ImmunoChem, Hamilton, Mont.), can be used inparenteral administration.

[0181] Adjuvants for mucosal administration include bacterial toxins,e.g., the cholera toxin (CT), the E. coli heat-labile toxin (LT), theClostridium difficile toxin A and the pertussis toxin (PT), orcombinations, subunits, toxoids, or mutants thereof. For example, apurified preparation of native cholera toxin subunit B (CTB) can be ofuse. Fragments, homologs, derivatives, and fusions to any of thesetoxins are also suitable, provided that they retain adjuvant activity.Preferably, a mutant having reduced toxicity is used. Suitable mutantsare described, e.g., in WO 95/17211 (Arg-7-Lys CT mutant), WO 96/6627(Arg-192-Gly LT mutant), and WO 95/34323 (Arg-9-Lys and Glu-129-Gly PTmutant). Additional LT mutants that can be used in the methods andcompositions of the invention include, e.g., Ser-63-Lys, Ala-69-Gly,Glu-110-Asp, and Glu-112-Asp mutants. Other adjuvants, such as abacterial monophosphoryl lipid A (MPLA) of, e.g., E. coli, Salmonellaminnesota, Salmonella typhimurium, or Shigella flexneri; saponins, orpolylactide glycolide (PLGA) microspheres, can also be used in mucosaladministration.

[0182] Adjuvants useful for both mucosal and parenteral administrationsinclude polyphosphazene (WO 95/2415), DC-chol (3 β-(N-(N′,N′-dimethylaminomethane)-carbamoyl) cholesterol; U.S. Pat. No. 5,283,185 and WO96/14831) and QS-21 (WO 88/9336).

[0183] Any pharmaceutical composition of the invention, containing apolynucleotide, a polypeptide, a polypeptide derivative, or an antibodyof the invention, can be manufactured in a conventional manner. Inparticular, it can be formulated with a pharmaceutically acceptablediluent or carrier, e.g., water or a saline solution such as phosphatebuffer saline, optionally complemented with a bicarbonate salt, such assodium bicarbonate, e.g., 0.1 to 0.5 M. Bicarbonate can beadvantageously added to compositions intended for oral or intragastricadministration. In general, a diluent or carrier can be selected on thebasis of the mode and route of administration, and standardpharmaceutical practice. Suitable pharmaceutical carriers or diluents,as well as pharmaceutical necessities for their use in pharmaceuticalformulations, are described in Remington's Pharmaceutical Sciences, astandard reference text in this field and in the USP/NF.

[0184] The invention also includes methods in which gastroduodenalinfections, such as Helicobacter infection, are treated by oraladministration of a Helicobacter polypeptide of the invention and amucosal adjuvant, in combination with an antibiotic, an antisecretoryagent, a bismuth salt, an antacid, sucralfate, or a combination thereof.Examples of such compounds that can be administered with the vaccineantigen and the adjuvant are antibiotics, including, e.g., macrolides,tetracyclines, β-lactams, aminoglycosides, quinolones, penicillins, andderivatives thereof (specific examples of antibiotics that can be usedin the invention include, e.g., amoxicillin, clarithromycin,tetracycline, metronidizole, erythromycin, cefuroxime, anderythromycin); antisecretory agents, including, e.g., H₂-receptorantagonists (e.g., cimetidine, ranitidine, famotidine, nizatidine, androxatidine), proton pump inhibitors (e.g., omeprazole, lansoprazole, andpantoprazole), prostaglandin analogs (e.g., misoprostil and enprostil),and anticholinergic agents (e.g., pirenzepine, telenzepine,carbenoxolone, and proglumide); and bismuth salts, including colloidalbismuth subcitrate, tripotassium dicitrate bismuthate, bismuthsubsalicylate, bicitropeptide, and pepto-bismol (see, e.g., Goodwin etal., Helicobacter pylori, Biology and Clinical Practice, CRC Press, BocaRaton, Fla., pp 366-395, 1993; Physicians' Desk Reference, 49^(th) edn.,Medical Economics Data Production Company, Montvale, N.J., 1995). Inaddition, compounds containing more than one of the above-listedcomponents coupled together, e.g., ranitidine coupled to bismuthsubcitrate, can be used. The invention also includes compositions forcarrying out these methods, i.e., compositions containing a Helicobacterantigen (or antigens) of the invention, an adjuvant, and one or more ofthe above-listed compounds, in a pharmaceutically acceptable carrier ordiluent.

[0185] Amounts of the above-listed compounds used in the methods andcompositions of the invention can readily be determined by those skilledin the art. In addition, one skilled in the art can readily designtreatment/immunization schedules. For example, the non-vaccinecomponents can be administered on days 1-14, and the vaccineantigen+adjuvant can be administered on days 7, 14, 21, and 28.

[0186] Methods and pharmaceutical compositions of the invention can beused to treat or prevent Helicobacter infections and, accordingly,gastroduodenal diseases associated with these infections, includingacute, chronic, and atrophic gastritis; and peptic ulcer diseases, e.g.,gastric and duodenal ulcers.

[0187] All twenty-four clones of the invention were isolated by atransposon shuttle mutagenesis method. Briefly, in this method, a TnMax9mini-blaM transposon was used for insertional mutagenesis of an H.pylori gene library established in E. coli. 192 E. coli clonesexpressing active β-lactamase fusion proteins were obtained, indicatingthat the corresponding target plasmids carry H. pylori genes encodingextracytoplasmic proteins. Individual mutants were transferred onto thechromosome of H. pylori P1 or P12 by natural transformation, resultingin 135 distinct H. pylori mutants. This method is described in furtherdetail, as follows.

[0188] The transposon TnMax9 (Kahrs et al., Gene 167:53, 1995) was usedto generate mutations in an H. pylori library in E. coli. As illustratedin FIG. 1A, TnMax9 contains, in addition to a cat_(GC)-resistance geneclose to the inverted repeat (IR), an unexpressed open reading frameencoding β-lactamase without a promoter or leader sequence (matureβ-lactamase, blaM; Kahrs et al., supra). For production ofextracytoplasmic BlaM fusion proteins resulting in ampicillin-resistant(amp^(R)) clones, expression of the cloned H. pylori genes in E. coli isobligatory. The minimal vector pMin2 (Kahrs et al., supra; see FIG. 1B),containing a weak constitutive promoter (P_(iga)) upstream of themultiple cloning site, was used for construction of the H. pylorilibrary to ensure expression of H. pylori genes in E. coli.

[0189] In construction of the library, H. pylori DNA was partiallydigested with Sau3A and HpaII, size fractionated by preparative agarosegel electrophoresis, and 3-6 kb fragments were ligated into the BglIIand ClaI sites of pMin2. The library was introduced into E. coli strainE181(pTnMax9), which is a derivative of HB101 containing the TnMax9transposon, by electroporation. This generated approximately 2,400independent transformants. More than 95% of the plasmids contained aninsert of between 3 and 6 kb, showing that the 1.7 Mb H. pylorichromosome was statistically covered. Since not every plasmid could beexpected to contain a target gene carrying an export signal, the librarywas partitioned into a total of 198 pools (24 pools of 20 clones and 174pools of 11 clones). Using a cotton swab, either eleven or twentyindividual colonies were inoculated in 0.5 ml LB medium in a eppendorftubes, vortexed, and 100 ml of the suspension was spread on LB agarplates supplemented with tetracycline and chloramphenicol to select formaintenance of both plasmids. Insertion of TnMax9 into the targetplasmids was induced with 100 mM isopropyl-b-D-thiogalactoside (IPTG)separately for each pool (Haas et al., Gene 130:23-21, 1993). Plasmidswere transferred into E145 by triparental mating, in which 25 ml of thedonor strain (E181), 25 ml of the mobilisator (KB101(pRK2013)), and 50ml of the recipient strain (E145) were mixed from correspondingbacterial suspensions (O.D.₅₅₀=10). The matings were performed for 2-3hours at 37° C. on nitrocellulose filters, which were placed on LBplates. Bacteria were suspended in 1 ml LB and aliquots were spread onLB plates containing chloramphenicol, tetracycline, and rifampicin. Eachpool gave rise to chloramphenicol-resistant transconjugates in E145,demonstrating that both transposition and conjugation were successful.Generally, several thousand chloramphenicol-resistant transconjugateswere obtained, but the number of amp^(R) colonies varied in differentpools, ranging from one to several hundred colonies. Two amp^(R)colonies from each positive pool were isolated, plasmid DNA wasextracted, and the DNA was characterized by further restrictionanalysis. Only those TnMax9 insertions of a single pool that mapped inobviously different plasmid clones, or in markedly different regions ofthe same clone, were used further.

[0190] From 158 of the 198 pools, ampicillin-resistant E145transconjugates were obtained (80%), showing that in several pools,TnMax9 inserted into expressed genes, resulting in production ofextracytoplasmic BlaM fusion proteins. Thus, a total of 192 amp^(R) E145clones could be isolated by conjugal transfer of plasmids from 198pools.

[0191] To analyze the mutant library, it was determined whether definedgene sequences inactivated by TnMax9 were represented once or severaltimes in the whole library. Five transposon-containing plasmidsconferring an amp^(R) phenotype to E145 (pMu7, pMu13, pMu75, pMu94, andpMu110) were randomly selected and DNA fragments flanking the TnMax9insert were isolated and used as probes in Southern hybridization of 120amp^(R) clones. The hybridization probes isolated from clones pMu7,pMu75, and pMu94 were between 0.9 and 1.1 kb in size, and hybridizedexclusively with the inserts of the homologous plasmids. In contrast,the TnMax9 flanking regions of clones pMu13 and pMu110 were 4.0 kb and5.5 kb, respectively. They each hybridized with the homologous plasmids,and with one additional clone of the library. Such a result wasexpected, since the chance of a probe to find a homologous sequence inthe library should be higher, the longer the hybridization probes.

[0192] In order to verify the insertion of the transposon into distinctORFs encoding putative exported proteins, the TnMax9-flanking DNA offive representative amp^(R) mutant clones (pMu7, pMu12, pMu18, pMu20,and pMu26) was sequenced, taking advantage of the M13 forward andreverse primers on TnMax9 (FIG. 1A). This analysis revealed that themini-transposon was inserted into different sequences in each plasmid,thereby interrupting ORFs encoding putative proteins. For two clones,the sequences located upstream of the blaM gene revealed a putativeribosome-binding site and a potential translational start codon (ATG).Other clones either revealed an ORF spanning the complete sequence(approximately 400 basepairs upstream and downstream of the TnMax9insertion) or termninating shortly after the site of TnMax9 insertion.The partial protein sequences from different ORFs were used for databasesearches, but no significant homologies with known proteins were found.

[0193] In a further approach, it was determined whether a known gene,like vacA, encoding the extracellular vacuolating cytotoxin of H.pylori, could be identified using this method and how often such amutation would be represented in the mutant library. A total celllysates of the 135 mutants were tested in an immunoblot using the H.pylori cytotoxin-specific rabbit antiserum AK197 (Schmitt et al., Mol.Microbiol. 12:307-319, 1994). Two mutants were identified, which nolonger produced the cytotoxin antigen (mutants P1-26 and P1-47) andpartial DNA sequencing of the insertion sites revealed that TnMax9 wasinserted at distinct positions in the vacA gene, 56 and 53 codonsdownstream of the ATG start codon, respectively.

[0194] Thus, the characterization of the mutant collection confirmedthat a representative gene library was constructed in E. coli, in whichtarget genes encoding exported H. pylori proteins were efficientlytagged by TnMax9.

[0195] In order to establish a collection of mutants lacking distinctexported proteins, the mutations had to be transferred back into the H.pylori chromosome. By means of natural transformation, 86 plasmids couldbe transformed into the original strain P1. H. pylori strains P1 or P12,which were naturally competent for DNA transformation, were transformedwith circular plasmid DNA (0.2-0.5 mg/transformation). Transformationsto streptomycin resistance were performed with chromosomal DNA (1mg/transformation), isolated from a streptomycin-resistant NCTC11637H.pylori mutant according to the procedure described in Haas et al. (Mol.Microbiol. 8:753-760). Selection was performed on serum platescontaining 4 mg/ml chloramphenicol or 500 mg/ml streptomycin. Thetransformation frequency for a given mutant was calculated as the numberof chloramphenicol-, streptomycin-, or erythromycin-resistant coloniesper cfu (average of three experiments). The blaM gene was deleted byNotI digestion, and the plasmid religated, in those plasmids that didnot transform strain P1 directly. This procedure, which resulted in atwenty to thirty-fold higher frequency of transformation, as compared tothe same plasmid containing blaM, resulted in 36 additional mutantsstrain P1. The blaM-deletion plasmids that still did not transformstrain P1 were used to transform the heterologous H. pylori strain P12,possessing an approximately 10-fold higher transformation frequencycompared to P1. This resulted in thirteen further mutants.

[0196] Thus, from the 192 amp^(R) plasmids a total of 135H. pylorimutants (122 mutants in P1 and 13 mutants in P12) were finally obtainedby selection on chloramphenicol resistance (70%). The transformationfrequency varied between different plasmids in the range of1×10⁻⁵-1×10⁻⁷. The remaining plasmids did not result in anytransformants. The collection was frozen as individual mutants in stockcultures at −70° C. To verify the correct insertion of themini-transposon into the H. pylori chromosome, ten representativemutants were tested by Southern hybridization of chromosomal DNA usingcat_(GC) DNA and the vector pMin2 as probes. Consistent with ourprevious experience concerning TnMax9-based shuttle mutagenesis of H.pylori, the mini-transposon was, in all cases, inserted into thechromosome without integration of the vector DNA, which probably meansby a double cross-over, rather than by a single cross-over event. Asjudged from the hybridization pattern obtained with the cat gene as aprobe, it appears that TnMax9 is located in different regions of thechromosome, showing that distinct target genes have been interrupted inindividual mutants.

[0197] The mutants were analyzed for motility, transformationcompetence, and adherence to KatoIII cells. Screening of the H. pylorimutant collection allowed identification of mutants impaired inmotility, natural transformation competence, and adherence to gastricepithelial cell lines. Motility mutants could be grouped into distinctclasses: (i) mutants lacking the major flagellin subunit FlaA and intactflagella; (ii) mutants with apparently normal flagella, but reducedmotility; and (iii) mutants with obviously normal flagella, butcompletely abolished motility. Two independent mutations, whichexhibited defects in natural competence for genetic transformation,mapped to different genetic loci. In addition, two independent mutantswere isolated by their failure to bind to the human gastric carcinomacell line KatoIII. Both mutants carried a transposon in the same gene,approximately 0.8 kb apart, and showed decrease autoagglutination, whencompared to the wild type strain.

[0198] The invention is further illustrated by the following examples.Example 1 describes isolation of DNA encoding a polypeptide of theinvention, HPO76. The methods described in Example 1 can be adapted forisolating nucleic acids encoding the other polypeptides of theinvention. Example 2 describes methods for obtaining the nucleic acidsof the invention from the deposited clones.

EXAMPLE 1 Preparation of Isolated DNA Encoding HPO76

[0199] 1.A. Preparation of Genomic DNA from Helicobacter Pylori

[0200]Helicobacter pylori strain ORV2001, stored in LB medium containing50% glycerol at −70° C., is grown on Colombia agar containing 7% sheepblood for 48 hours under microaerophilic conditions (8-10% CO₂, 5-7% O₂,85-87% N₂). Cells are harvested, washed with phosphate buffer saline(PBS; pH 7.2), and DNA is then extracted using the Rapid Prep GenomicDNA Isolation kit (Pharmacia Biotech).

[0201] 1.B. PCR Amplification

[0202] The DNA fragment is amplified from genomic DNA, as preparedabove, by the Polymerase Chain Reaction (PCR) using the followingprimers: -N-terminal primer: 5′-GCC[GAGCTC]ITATCGTATGGACTTAGAACAT-3′(SEQ ID NO:145) -C-terminal primer:5′-GCC[CTCGAG]ATTAGAATAAGTGTTGTTTAAAATC-3′. (SEQ ID NO:146)

[0203] Both primers include a clamp (GCC) and a restriction enzymerecognition sequence for cloning purposes (SacI (GAGCTC) and XhoI(CTCGAG) recognition sequences). The underlined sequences in bothprimers represent clone 76-specific sequences. The N-terminal primer isdesigned so that the amplified product does not encode the leadersequence and the potential cleavage site.

[0204] Amplification of gene-specific DNA is carried out using Pwo DNAPolymerase (Boehringer Mannheim), which is a proof-reading polymerase,according to general guidance provided by the manufacturer. Because ofthe exonuclease activity of the polymerase, two reaction mixtures(mixtures 1 and 2) are first prepared separately and combined just priorto amplification. These mixtures are as follows: Ingredient (finalconc.) Mixture 1 (l) Mixture 2 (l) distilled H₂O 160 79 dNTPs (200 Meach) 40 — 10x PCR buffer — 20 primers (100 nM each) 1 — DNA template(200 ng) 2 — as obtained in 1.A.

[0205] Amplification is carried out as follows: Number of Cyclingconditions Temp. (° C.) Time (min.) cycles Initial denaturing 96 4  1step Denaturing step 94   0.5 20 Annealing step 50 1 20 Extension step72 1 20 Final extension step 72 5  1

[0206] 1.C. Transformation and Selection of Transformants

[0207] A single PCR product of 522 basepairs is thus amplified and isthen digested at 37° C. for 2 hours with SacI and XhoI concurrently in a20 μl reaction volume. The digested product is ligated to similarlycleaved pET28a (Novagen) that is dephosphorylated prior to the ligationby treatment with Calf Intestinal Alkaline Phosphatase (CIP). The genefusion constructed in this manner allows one-step affinity purificationof the resulting fusion protein because of the presence of histidineresidues at the N-terminus of the fusion protein, which are encoded bythe vector.

[0208] The ligation reaction (20 μl) is carried out at 14° C. overnightand then is used to transform 100 μl fresh E. coli XL1-blue competentcells (Novagen). The cells are incubated on ice for 2 hours, thenheat-shocked at 42° C. for 30 seconds, and returned to ice for 90seconds. The samples are then added to 1 ml LB broth in the absence ofselection and grown at 37° C. for 2 hours. The cells are then plated outon LB agar plus kanamycin (50 μg/ml final concentration) at a 10× andneat dilution and incubated overnight at 37° C. The following day, 50colonies are picked onto secondary plates and incubated at 37° C.overnight.

[0209] Five colonies are picked into 3 ml LB broth supplemented withkanamycin (100 μg/ml) and grown overnight at 37° C. Plasmid DNA isextracted using the Quiagen mini-prep. method and quantitated by agarosegel electrophoresis.

[0210] PCR is performed with the gene-specific primers under theconditions stated above and transformant DNA is confirmed to contain thedesired insert.

[0211] If PCR-positive, one of the five plasmid DNA samples (500 ng)extracted from the E. coli XL1-blue cells is used to transform competentBL21 (IDE3) E. coli competent cells (Novagen; as described previously).Transformants (10) are picked onto selective kanamycin (50 μg/ml)containing LB agar plates and stored as a research stock in LBcontaining 50% glycerol.

[0212] 1.D. Recombinant Production of the Protein

[0213] Frozen stock (10 μl) is plated onto selection plates and grownfor single colonies overnight at 37° C. A few cells are harvested fromthe plate and used as the inoculum for an overnight starter culture (3ml) at 37° C. The following day, a sample (time ‘t’=0) is collected andcentrifuged at 14,000 rpm for 3 minutes (samples are standardized byOD₆₀₀ for each time-point). The supernatant is discarded and the cellsare stored at −20° C. for SDS-PAGE. This allows detection of leakyexpression in the absence of the inducer IPTG. The overnight starterculture is then used to inoculate LB medium containing kanamycin (100μg/ml) at a dilution of 1:50 (starting OD₆₀₀=0.05−0.1). The cells aregrown to an OD₆₀₀ of 1.0, a sample is harvested for SDS-PAGE(pre-induction sample), and the remaining culture is induced with 1 mMIPTG. The cultures are grown for 4 hours and samples are taken everyhour.

[0214] The culture is spun in a centrifuge at 6000× g for 20 minutes at4° C. The supernatant is discarded and the pellets are resuspended in 50ml of cold 50 mM Tris-HCl (pH 8.0), 2 mM EDTA, and spun as is describedabove. The supernatant is discarded and the cells are stored at −70° C.

[0215] 1.E. Protein Purification

[0216] Pellets obtained from a 1 liter culture prepared as described in1.D. are thawed and resuspended in 20 ml of ice cold 20 mM Tris-HCl (pH8.0), 0.5 M NaCl, 5 mM Imidazole. Lysozyme is added to a concentrationof 0.1 mg/ml and the suspension is homogenized using a high speedhomogenizer (Turrax), and subsequently is treated in a sonicator(Branson, Sonifier 450). To remove DNA, Benzonase (Merck) is used at afinal concentration of 1 U/ml. The suspension is centrifuged at 40,000×g for 20 minutes and the supernatant is filtered through a 0.45 μmmembrane. The supernatant is loaded onto an IMAC column (12 ml of resin)that has been prepared by immobilizing Ni⁺⁺ according to therecommendations of the manufacturer (Pharmacia). The column is washedwith 10 column volumes of 20 mM Tris-HCl (pH 8.0), 0.5 M NaCl, 60 mMImidazole. The recombinant protein is eluted with 6 volumes of 20 mMTris-HCl (pH 7.9), 0.5 M NaCl, 500 mM Imidazole, 0.1% Zwittergent 3-14.

[0217] The elution profile is monitored by measuring the absorbance ofthe fractions at OD 280 nm. An aliquot of each fraction is analyzed onSDS-PAGE gels and stained with Coomassie blue (Phast System—Pharmacia),and the fractions corresponding to the protein peak are then pooled andconcentrated. To remove elution buffer, the fraction is passed over aG25 Sephadex column (Pharmacia), equilibrated in PBS (pH 7.4). Theprotein solution is filter-sterilized through a 0.45 μm membrane, andthe protein concentration is determined by the BCA micromethod (Pierce).The protein solution is stored at −70° C.

[0218] 1.F. Evaluation of the Protective Activity of the PurifiedProtein

[0219] Groups of 8 Swiss-Webster mice (Taconic) are immunized orallywith 25 μg of the purified recombinant protein, admixed with 5 μg ofcholera toxin (Calbiochem) in physiological buffer. Mice are immunizedon days 0, 7, 14, and 21. Fourteen days after the last immunization, themice are challenged with H. pylori strain ORV2001 grown in liquid media(the cells are grown on agar plates as described in 1.1. and, afterharvest, the cells are resuspended in Brucella broth; the flasks areincubated overnight at 37° C.). Fourteen days after challenge, the miceare sacrificed and their stomachs are removed. The amount of H. pyloriis determined by measuring the urease activity in the stomach and byculture.

[0220] 1.G. Production of Monospecific Polyclonal Antibodies

[0221] 1.G.1. Hyperimmune Rabbit Antiserum

[0222] New Zealand rabbits are injected both subcutaneously andintramuscularly with 100 μg (in total) of the purified fusionpolypeptide as obtained in 1.E., in the presence of Freund's completeadjuvant in a total volume of approximately 2 ml. Twenty-one and 42 daysafter the initial injection, booster doses, which are identical topriming doses, except that Freund's incomplete adjuvant is used, areadministered in the same way. Fifteen days after the last injection,animal serum is recovered, decomplemented, and filtered through a 0.45μm membrane.

[0223] 1.G.2. Mouse Hyperimmune Ascitic Fluid

[0224] Ten mice are injected subcutaneously with 10-50 μg of thepurified fusion polypeptide as obtained in 1.E., in the presence ofFreund's complete adjuvant in a volume of approximately 200 μl. 7 and 14days after the initial injection, booster doses, which are identical topriming doses, except that Freund's incomplete adjuvant is used, areadministered in the same way. 21 and 28 days after the initialinfection, mice receive 50 μg of the antigen alone intraperitoneally. Onday 21, mice are also injected intraperitoneally with sarcoma 180/TGcells CM26684 (Lennette et al., Diagnostic procedures for viral,rickettsial, and chlamydial infections, (1979) 5^(th) Ed. WashingtonD.C., American Public Health Association). Ascites are collected 10-13days after the last injection.

[0225] 1.H. Purification by Immunoaffinity

[0226] 1.H.1. Purification of Specific IgGs

[0227] An immune serum as prepared in section 1.G. is applied to aprotein A Sepharose 4 Fast Flow column (Pharmacia) equilibrated in 100mM Tris-HCl (pH 8.0). The resin is washed by applying 10 column volumesof 100 mM Tris-HCl and 10 volumes of 10 mM Tris-HCl (pH 8.0) to thecolumn. IgGs are eluted with a 0.1 M glycine buffer (pH 3.0) and arecollected as 5 ml fractions to which is added 0.25 ml 1 M Tris-HCl (pH8.0). The optical density of the eluate is measured at 280 nm and thefractions containing the IgGs are pooled, and, if necessary, storedfrozen at −70° C.

[0228] 1.H.2. Preparation of the Column

[0229] An appropriate amount of CNBr-activated Sepharose 4B gel (1 g ofdried gel provides for approximately 3.5 ml of hydrated gel; gelcapacity is of from 5 to 10 mg coupled IgGs per ml of gel) manufacturedby Pharmacia (17-0430-01) is suspended in 1 mM HCl buffer and washedwith a buchner by adding small quantities of 1 mM HCl buffer. The totalvolume of buffer is 200 ml per gram of gel.

[0230] Purified IgGs are dialyzed for 4 hours at 20±5° C. against 50volumes of 500 mM sodium phosphate buffer (pH 7.5). Then they arediluted in 500 mM phosphate buffer (pH 7.5) to a final concentration of3 mg/ml.

[0231] IgGs are incubated with the gel overnight at 5±3° C., understirring. The gel is packed into a chromatography column and washed with2 column volumes of 500 mM phosphate buffer (pH 7.5), then 1 volume of50 mM sodium phosphate buffer, 500 mM NaCl (pH 7.5). The gel is thentransferred to a tube and further incubated in 100 mM ethanolamine, (pH7.5) for 4 hours at room temperature under stirring, then washed twicewith 2 column volumes of PBS. The gel is then stored in 1/10,000 PBSmerthiolate. The amount of IgGs coupled to the gel is determined bymeasuring the optical density (OD) at 280 nm of the IgG solution and thedirect eluate, plus washings.

[0232] 1.H.3. Adsorption and Elution of the Antigen

[0233] An antigen solution in 50 mM Tris-HCl (pH 8.0), 2 mM EDTA, forexample, the supernatant obtained in 1.E. after the Benzonase treatment,centrifugation, and filtration through a 0.45 μm membrane, is applied toa column equilibrated with 50 mM Tris-HCl (pH 8.0), 2 mM EDTA, at a flowrate of about 10 ml/hour. Then the column is washed with 20 volumes 50mM Tris-HCl (pH 8.0), 2 mM EDTA. Alternatively, adsorption can beachieved in a batch that is let to stand overnight at 5±3° C., understirring.

[0234] The gel is washed with 2 to 6 volumes of 10 mM sodium phosphatebuffer (pH 6.8). The antigen is eluted with 100 mM glycine buffer (pH2.5). The eluate is recovered in 3 ml fractions to which is added 150 μl1 M sodium phosphate buffer (pH 8.0). OD is measured at 280 nm for eachfraction; those containing the antigen are pooled and stored at −20° C.

EXAMPLE 2 Preparation of Isolated DNA Encoding the Polypeptides of theInvention from the Deposited Clones.

[0235] As mentioned above, E. coli strains including plasmids containingnucleic acids encoding HPO76 (98197), HPO18 (98210), HPO121(98201),HPO45 (98208), HPO101(98198), HPO116 (98200), HPO7 (98211), HPO104(98199), HPO15 (98214), HPO58 (98206), HPO132 (98202), HPO9 (98203),HPO38 (98204), HPO87 (98205), HPO71(98217), HPO70 (98219), HPO80(98215), HPO95 (98216), HPO98 (98218), HPO57 (98220), HPO50 (98207),HPO64 (98213), HPO54 (98212), and HPO42 (98209) were deposited in E.coli strain DH5α under the Budapest Treaty with the American TypeCulture Collection (ATCC; Rockville, Md.) on Oct. 9, 1996 and weredesignated with accession numbers indicated in parentheses above. Theseplasmids each contain a genomic DNA BglII-ClaI insert from H. pyloristrain P1 or P12 (referred to as 69-A and 888-0 in Haas et al., Mol.Microbiol. 8:753, 1993). Each of the inserts are disrupted by thepresence of transposon TnMax9 (Kahrs et al., Gene 167:53, 1995). DNAmolecules lacking the transposon can be amplified from the plasmidsusing standard PCR techniques, such as inverse and recombinant PCR (see,e.g., Innis et al., supra), so that a full-length H. pylori insert isreconstituted. For example, the H. pylori sequences flanking thetransposon can each be amplified by PCR, and then ligated together toform the full-length H. pylori gene lacking the transposon. Primers thatcan be used in these methods for each of the twenty-four clones of theinvention are shown in Table 1.

EXAMPLE 3 Purification of Recombinant H. Pylori Antigen from Clone 76(HPO76)

[0236] A pellet of E. coli expressing HPO76 is homogenized in 5 mMimidazole, 500 mM sodium chloride, 20 mM Tris-HCl (pH 7.9) bymicrofluidization at a pressure of 15,000 psi, and clarified bycentrifugation at 4000-5000 g.

[0237] Method 1

[0238] The pellet containing cloned protein is suspended in buffercontaining 2% N-octyl glucoside (NOG) and is homogenized. The NOGsoluble protein is removed by centrifugation. The pellet is extractedone more time with 2% NOG. After centrifugation, the pellet is dissolvedin 8 M urea. The urea-solubilized protein is diluted with an equalvolume of 2 M arginine and dialyzed against 1 M arginine for 24-48 hoursto remove urea. The cloned protein remains in solution. SDS-PAGE andCoomassie staining, followed by densitometric scanning, shows that theprotein is 80-85% pure cloned antigen.

[0239] Method 2

[0240] The pellet containing cloned protein is solubilized in 6 Mguanidine hydrochloride and is passed through an IMAC column chargedwith Ni⁺⁺. The bound antigen is eluted with 8 M urea (pH 8.5).β-mercaptoethanol is added to eluted protein to a final concentration of1 mM, then passed through a Sephadex G-25 column equilibrated in 0.1 Macetic acid. Protein eluted from Sephadex G-25 column is slowly added to4 volumes of 50 mM phosphate (pH 7.0). The protein remains in solution.

[0241] Purification of Recombinant Proteins

[0242] Recombinant proteins expressed as Histidine-tagged fusionproteins can be solubilized and purified by using a metal affinitycolumn (nickel column). The bound protein can be eluted with imidazolebuffer, with or without urea, or by using low pH buffers, with orwithout urea. Urea or guanidine hydrochloride-denatured proteins canthen be renatured using appropriate renaturing buffers. With a number ofrecombinant H. pylori antigens (HpaA and clone 76), renaturationconditions using arginine hydrochloride (0.25-1 M) have been determined.

[0243] Recombinant proteins without a His-tag can be solubilized andpurified using immunoaffinity, ion-exchange, sizing, and/or hydrophobicchromatography. Proteins expressed as insoluble aggregates in inclusionbodies can be solubilized in denaturing agents, such as 8 M urea or 6 Mguanidine hydrochloride. Appropriate folding and renaturation canreadily be determined by one skilled in the art.

[0244] The above pellet containing cloned protein is suspended in 50 mMNaPO₄ (pH 7.5) containing 1% weight/volume N-octyl glucoside (NOG) andmixed vigorously. The NOG soluble impurities are removed bycentrifugation. The remaining pellet is extracted one more time with the1% NOG solution to further remove impurities. After centrifugation, thepellet is solubilized in 8 M urea, 50 mM Tris (pH 8.0). The Ureasolubilized protein is diluted with an equal volume of 2 M Arginine, 50mM Tris (pH 8.0), and is dialyzed against 1 M Arginine, 50 mM Tris, 50mM NaCl (pH 8.0) for 24-48 hours to remove urea. The cloned proteinremains in solution following dialysis. SDS-PAGE and Coomassie stainingfollowed by densitometric scanning shows that the protein is 80-85% purecloned antigen.

EXAMPLE 4 Method for Production of Transcriptional Fusions LackingHis-Tags

[0245] Methods for amplification and cloning of DNA encoding HPO76 as atranscriptional fusion lacking His-tags are described as follows. Thesemethods can readily be adapted by one skilled in the art for similaramplification and cloning of DNA encoding the other polypeptides of theinvention.

[0246] Amplification of Clone 76 DNA

[0247] Design of PCR Primers for Cloning

[0248] Two PCR primers are designed based on the complete gene sequence(see table 1).

[0249] The N-terminal primer (FC1) is designed to include the ribosomebinding site of the target gene (underlined), the ATG start site (bold),and the leader sequence (with cleavage site). It includes a clamp (GCC)at the 5′ most end, and a SacI recognition sequence (GAGCTC) for cloningpurposes.

[0250] The C-terminal primer (RN2) includes an XhoI recognition sequencefor cloning purposes, and the natural TAA stop codon (bold). N-terminalprimer (FC1) 5′GCC[GAGCTC]CAAGCAAAAAAATGTCAATTAAAAGGG3′ (SEQ ID NO:)C-terminal primer (RN2) 5′GCC[CTCGAG]GTCTAAATTAGAATAAGTGTTGTT 3′ (SEQ IDNO:)

[0251] Amplification of each specified gene can be achieved by employingFC1/RN2 primers for any of the genes described (see Table 1).

[0252] PCR Conditions

[0253] Amplification of gene-specific DNA is carried out using Pwo DNAPolymerase (Boehringer Mannheim) under the following conditions. Due tothe exonuclease activity of the polymerase, two reaction mixtures areprepared separately and combined just prior to amplification. Reactioningredients: Ingredient (final conc.) Mixture 1 (μl) Mixture 2 (μl)distilled H₂O 160 79 dNTPs (200 μM each) 40 — 10X buffer — 20 primer 1(100 nM) 1 — primer 2 (100 nM) 1 — Template (200 ng) 2 0 Cyclingcondition Temp (° C.) Time (min.) Number of cycles Initial denaturingstep 96 4  1 Denaturing step 94   0.5 20 Annealing step 50 1 20Extension step 72 1 20 Final extension step 72 1  1

[0254] A single PCR product of 624 basepairs is amplified and clonedinto SacI-XhoI cleaved pET 24, allowing construction of atranscriptional fusion and expression of HPO76 antigen in the absence ofa His-tag. In this instance, expressed product can be purified as adenatured protein that is re-folded by dialysis into 1 M arginine.

[0255] Cloning into pET 24 allows transcription from the T7 promoter,supplied by the vector, but relies upon binding of the RNA-specific DNApolymerase to the intrinsic ribosome binding site for HPO76, and therebyexpression of the complete ORF. The amplification, restriction, andcloning protocols are as previously described for constructingtranslational fusions. TABLE 1 RE-CONSTRUCTION OF A COMPLETE ORF BYRECOMBINANT PCR F′ denotes forward primer R′ denotes reverse primerC′ denotes coding strand N′ denotes non-coding strand Alt FC1 and RN2primers have incorporated at their 5′ end a clamp and a recognitionsequence for cloning purposes GGC clamp present for amplification andcloning of entire gene sequence from chromosomal DNA [X] denotes anynucleotide sequence not present in the completed gene sequence ()Identifies region of overlap between the two original PCR products, andis consistently 10 nucleotides long for each clone Length CLONE No.Prtmer type nt positions Primer sequence (5′-3′) of gene seq. Tm (oC) 76FC1 304-330 GCC[x] CAAGCAAAAAAATGTCAATTAAAAGGG 27 70 RN1 413-391TAAGTCCATACGATAGCCTATG 22 62 FC2 404-436 (TATGGAACTTA)GAACATTTTAACACGCTCTATTA 33 60 RN2 927-904 GCC[X] GTCTAAATTAGAATAAGTGTTGTT 24 60 18 FC1 101-124GCC[X] AATATATGGGAACTTAATGAGAAT 24 60 RN1 227-206 TGCGAGATTTAACCTGTTTTCA22 60 FC2 218-249 (AAATCTCGCA) GAAATCTTTCACAAGCGAGCAA 32 60 RN2 922-901GCC [X] ATGTCATGTCAAACTATGAAGC 22 60 121 FC1 141-164 GCC[X] TCACAATGGATAAAAACAACAACA 24 62 RN1 451-473 GCCCTTTTGTTTAGGGGTTAG 2162 FC2 455-485 (ACAAAAGGGC) TTTTTAGAGCATGTGAGCCATC 32 62 RN2 814-796 GCC[X] CTGTCCAAATCAGCCACCC 19 60 45 FC1  1-26 GCC[X] ATGAAAAGATTTGATTTGTTTTTATC 26 62 RN1 299-278 AAGCCGTATTGTTTGTTTTGGC22 62 FC2 290-323 (AATACGGCTTTAAAGCTATAGAAAATTTAAACGC) 34 60 RN2 603-582GCC[X] TTAAATATCCCAATCCTGCCAC 22 62 101 FC1 308-332GCC[X] GAAGGATTTATTATGATTAAAAGAA 25 60 RN1 497-474AACCTAATTTGAAATTCAAACCAT 24 60 FC2 488-519 (AAATTAGGTT)TTGTAGGCTTTGCCAATAAATG 32 60 RN2 893-869GCC[X] AAGGAATAAATTAGAAAGTGAAGAA 25 62 116 FC1 236-259 GCC[X] CGCATTGATTTGATGAATAAACC 23 62 RN1 434-416 CGCCTATAACCGCTCCATT 19 60FC2 425-456 (GTTATAGGCG) ATAAAGGTTTAACGCAGCTAAG 32 60 RN2 812-790 GCC[X] CTCACTAAAAAGCAATTTTTGAG 23 60 7 FC1 195-220 GCC[X] TAAGGAATGAAGTTGATAAAATTTGT 26 64 RN1 349-327 GCATTTTCATTCATTCTTTGGAC23 60 FC2 339-371 (ATGAAAATGC) ACGCCCAAATAATAAGGAAGTA 32 60 RN2 738-717GCC [X] GGATTTATTGAGCTTTCCCCTT 22 62 104 FC1 251-271 GCC[X] AAAGGGCGAAAATGAGCAAGA 21 60 RN1 429-407 TAAAATAACCAACAGAGTGATCA 2360 FC2 420-452 (GGTTATTTTA) GTGGATATTTGGGTTTATAGCGA 33 62 RN2 784-761GCC [X] TTTTTTAAGAATCACTTTCTTCGG 24 62 58 GC1 118-143 GCC[X] ATAGGAACAAGCATGTTTTTTAAAAC 26 66 RN1 434-413 TGAAGTCTTGCGATTTTTGCTT22 60 FC2 425-454 (CAAGACTTCA) AAAAAGAAGGAGCGGTTGCC 30 60 RN2 650-630GCC [X] CTGGCTTATTGCGTATCATC 20 60 132 FC1 294-314 GGC[X] GGAAGAATAATGCTCGCTTCC 21 62 RN1 409-378 ACTGGAGTGTGGATAAAACTAT 22 60FC2 400-430 (ACACTCCAGT) AGATGCTTTCCCGGATATTTC 31 60 RN2 761-741 GCC[X] CTATTCTCCAGGGATATGGCC 21 64 9 FC1 211-233 GCC[X] GATGGATTTTTTATGGGGGTGAG 23 64 RN1 347-328 GGCACTGCCGCAGATTCTA 19 60FC2 338-370 (CGGCAGTGCC) TTTAGCCTATTATTTAGAAGCGA 33 60 RN2 686-665 GCC[X] ATGGTATTTGTCTAAGACCCTC 22 62 38 FC1 220-242 GCC[X] AAAAGGGTTTTAAATAATGGCTG 23 60 RN1 348-327 ACAAGGATAAAAAACGCGCTAA 2260 FC2 239-371 (TTATCCTTGT) TGCTGGCTTGGTTTTTTTTAATT 33 60 RN2 597-575GCC [X] AAGATTCTAAAAGGGCTTCAAAT 23 60 71 FC1  1-25 GCC[X] ATGTTGAAATTTAAATATGGTTTGA 25 60 RN1 274-254 AAACCCCACTCTTATCATCGG 2162 FC2 265-294 (AGTGGGGTTT) TTTTAGGGGGTGGGTATGCT 30 60 RN2 524-505 GCC[X] GAGCCTACAGGTTGCTTGC 20 60 70 FC1  1-23 GCC[X] ATGGTATTTGACAGAACAATCAG 23 62 RN1 115-96  GAAAAGCCACCCCGCTTATT 20 60FC2 106-137 (GTGGCTTTTC) AAAAAGAGTGGGTGCAACAATT 32 60 RN2 495-471 GCC[X] TTAGGAATAGCATAACAAACAAACG 25 66 80 FC1  1-25 GCC[X] ATGTTAGAAAAATTGATTGAAAGAG 25 62 RN1 106-95  TGAACACATAGCCTAAAACCAC21 62 FC2  97-127 (TATGTGTTCA) TGAAAGAGTTGTGGCACATGC 31 62 RN2 435-415GCC [X] TTATGCGATAGGGGGCGTATC 21 66 95 FC1  1-27 GCC[X] ATGAAAAAATTTTTTTCTCAATCTTT 27 60 RN1 64-46 TGGCCAGTAGCGCGTTCAT 19 60FC2 55-98 (CTACTGGCCA) TGGATGGCAATGGCGTTTTTTTAG 34 68 RN2 432-408 GCC[X] TTATTGATGAACATTAACCATTAAA 25 60 98 FC1  1-22 GCC[X] ATGAAAACCTTTAAAAACCTGC 22 58 RN1 43-23 TAGCGATCAGGCTAAAACAGA 21 60FC2 34-62 (CTGATCGCTA) TGAGTTGGCTCCAAGCGGA 29 60 RN2 336-313 GCC[X] TTAAAACTCATAGCGTTTTTCAAT 24 60 42 FC1 18-51 GCC[X] GAGAGTAGTGGCAGAGTTTATGCTGATTCCC 34 98 RN1 380-351(AACTTTTC)TCTATCCCAATTCGTTACGCTC 30 64 FC2 366-396(GGATAGA)GAAAAGTTTGGCGTCAAAAGTTGG 31 68 RN2 822-801 GCC[X] GGCTTAAACTGGAACGGATTTC 22 64 50 FC1 140-170 GCC[X] TAAAGTTTGCTAAAAAGATGGTTTTAATTTC 31 76 RN1 297-270(GACTTCTAAAG)CGTCCTTTTTTTCTTTA 28 56 FC2 287-314(CTTTA)GAAGTCATTAAACAAAGAGGGGT 29 64 RN2 607-584 GCC[X] CCCATCTTTAGAAATCAACCCCCA 24 70 64 FC1 23-50 GCC[X] GAAATAAGGAGTTTGTATGCAACAGCG 28 80 RN1 225-149(A)AGCTTTTCATTATCTTCCCCATAAGC 27 74 FC2 216-244(TGAAAAGCT)TTTAGCGAAGCGATCAAGCC 29 60 RN2 1039-1012 GCC[X] CCCAATACTTTTATTGATTCACCATTTC 28 74 54 FC1 21-48 GCC[X] CAATAAAACACCAAAATGAATGAGTTAC 28 68 RN1 352-327(A)GATTTTGTTTTGAGCGTTAGAAATG 26 66 FC2 345-376(CAAAATC)TATAAACTCAATCAAGTCAAAAATG 32 62 RN2 1280-1255 GCC[X] GCATTTACCCCCTAAAAACTATAAAC 26 70 15 FC1 14-35 GCC[X] CTGAAGGGTGTATGGTATTAGG 22 64 RN1 157-132(C)ACCATACATGTATCCTGCATTAATG 26 68 FC2 147-179(CATGTATGGT)GTAGCAAAGAATTTTAAGGAGGC 33 64 RN2 377-349 GCC[X] CGTTAAAACTAAAGTTCTATTTTTAATTC 29 70 57 FC1 13-39 GCC[X] GTAAGGAATGAGATGATAAAGAGTTGG 27 74 RN1 267-244(T)GGAATATTCTGATCCACGCCATC 24 68 FC2 258-294(GAATATTCC)AAAAGCCGTTTTTTATTACAGAAGAGC 37 76 RN2 957-934 GCC[X] CTAAACTCTGGCTTATTGCGTATC 24 68 87 FC1  1-22 GCC[X] ATGCGTTTATTATTGTGGTGGG 22 62 RN1 27-3  (C)AATACCCACCACAATAATAAACGCAT25 66 FC2 18-50 (GTGGGTATT)GGTATTATCGCTCTTTTTAAATCC 33 64 RN2 519-498GCC [X] TTAAATTTTTAGGGAAAGGGTA 22 62 CONDITIONS FOR RECOMBINANT PCR Twoindependent PCR reactions are carried out for FC1/RN1 and fC2/RN2primers under the same conditions proposed for cloning genes forexpression. After 20 cycles, the product of each reaction is used astemplate for a further 20 cycles with FC1/RN2 only The product willencompass the full length gene minus the transposon. The presence ofrestriction sites at the 5′ ends of these primers allows forcloning/expression studies.

[0256]

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 146 <210> SEQ ID NO 1<211> LENGTH: 982 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (320)...(880) <221>NAME/KEY: sig_peptide <222> LOCATION: (320)...(400) <221> NAME/KEY:mat_peptide <222> LOCATION: (401)...(880) <400> SEQUENCE: 1 agattagcagcagcagggat ttttaaattc ctggccaaca ggggcggttg gaaaaaaata 60 cgattaaaaaggcaaacgct ttgaaagtat tttttcatag aaattccctt ttgttaaatg 120 attgaagttggtgattatac ctatttgtat cttaaaaatt tgattttaaa agtttgagat 180 ggttttgtaggtgtatccca cttatccaat ttatatcaat attttcactc taaaaccctc 240 atccttgataaaaaattaaa ccttttagaa aaataaccga ttttagggtg taactttaat 300 tcaacaagaaggatttatt atg att aaa aga att gct tgt att tta agc ttg 352 Met Ile LysArg Ile Ala Cys Ile Leu Ser Leu -25 -20 agt gcg agt tta gcg ctg gct ggcgaa gtg aat ggg ttt ttc atg ggt 400 Ser Ala Ser Leu Ala Leu Ala Gly GluVal Asn Gly Phe Phe Met Gly -15 -10 -5 gcg ggt tat cag caa ggt cgt tatggt cct tat aac agc aat tac tct 448 Ala Gly Tyr Gln Gln Gly Arg Tyr GlyPro Tyr Asn Ser Asn Tyr Ser 1 5 10 15 gat tgg cgc cat ggc aat gat ctttat ggt ttg aat ttc aaa tta ggt 496 Asp Trp Arg His Gly Asn Asp Leu TyrGly Leu Asn Phe Lys Leu Gly 20 25 30 ttt gta ggc ttt gcc aat aaa tgg tttggg gct agg gtg tat ggc ttt 544 Phe Val Gly Phe Ala Asn Lys Trp Phe GlyAla Arg Val Tyr Gly Phe 35 40 45 tta gat tgg ttt aac act tca ggg aca gaacac acc aaa acc aat ttg 592 Leu Asp Trp Phe Asn Thr Ser Gly Thr Glu HisThr Lys Thr Asn Leu 50 55 60 ctc acc tat ggt ggc ggt ggc gat ttg att gtcaat ctc att cct ttg 640 Leu Thr Tyr Gly Gly Gly Gly Asp Leu Ile Val AsnLeu Ile Pro Leu 65 70 75 80 gat aaa ttc gct cta ggt ctc atc ggt ggc gttcaa tta gcc gga aac 688 Asp Lys Phe Ala Leu Gly Leu Ile Gly Gly Val GlnLeu Ala Gly Asn 85 90 95 act tgg atg ttc cct tat gat gtc aat caa acg agattc cag ttc tta 736 Thr Trp Met Phe Pro Tyr Asp Val Asn Gln Thr Arg PheGln Phe Leu 100 105 110 tgg aat tta ggc gga aga atg cgt gtt ggg gat cgcagt gcg ttt gaa 784 Trp Asn Leu Gly Gly Arg Met Arg Val Gly Asp Arg SerAla Phe Glu 115 120 125 gca ggc gtg aaa ttc cct atg gtt aat caa ggc aacaaa gat gtt agg 832 Ala Gly Val Lys Phe Pro Met Val Asn Gln Gly Asn LysAsp Val Arg 130 135 140 gct tat ccg cta cta ttc ttg ggt atg tgg att atgttc ttc act ttc 880 Ala Tyr Pro Leu Leu Phe Leu Gly Met Trp Ile Met PhePhe Thr Phe 145 150 155 160 taatttattc ctttcattcg ctcttcttca tcaaatcaaccctaacccac tcttaaaagg 940 ttggggttca aaaatctttt tcataaataa aatttgcctt aa982 <210> SEQ ID NO 2 <211> LENGTH: 187 <212> TYPE: PRT <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 2 Met Ile Lys Arg Ile Ala Cys IleLeu Ser Leu Ser Ala Ser Leu Ala 1 5 10 15 Leu Ala Gly Glu Val Asn GlyPhe Phe Met Gly Ala Gly Tyr Gln Gln 20 25 30 Gly Arg Tyr Gly Pro Tyr AsnSer Asn Tyr Ser Asp Trp Arg His Gly 35 40 45 Asn Asp Leu Tyr Gly Leu AsnPhe Lys Leu Gly Phe Val Gly Phe Ala 50 55 60 Asn Lys Trp Phe Gly Ala ArgVal Tyr Gly Phe Leu Asp Trp Phe Asn 65 70 75 80 Thr Ser Gly Thr Glu HisThr Lys Thr Asn Leu Leu Thr Tyr Gly Gly 85 90 95 Gly Gly Asp Leu Ile ValAsn Leu Ile Pro Leu Asp Lys Phe Ala Leu 100 105 110 Gly Leu Ile Gly GlyVal Gln Leu Ala Gly Asn Thr Trp Met Phe Pro 115 120 125 Tyr Asp Val AsnGln Thr Arg Phe Gln Phe Leu Trp Asn Leu Gly Gly 130 135 140 Arg Met ArgVal Gly Asp Arg Ser Ala Phe Glu Ala Gly Val Lys Phe 145 150 155 160 ProMet Val Asn Gln Gly Asn Lys Asp Val Arg Ala Tyr Pro Leu Leu 165 170 175Phe Leu Gly Met Trp Ile Met Phe Phe Thr Phe 180 185 <210> SEQ ID NO 3<211> LENGTH: 843 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (262)...(777) <400>SEQUENCE: 3 ccaatggagg cgtttccaaa aacccaaacg ggcgcttttt aaagaaaaatctcaaaaaat 60 tcagggagca agcggtaaaa atcgtagaaa aacgcttgat aaaagagaatatgcaactga 120 gcgattttaa tgaagaagaa ttaaaaatca tgtttgaagc tgaagaaaaaaggttgttag 180 agcaaatcca cgctaaagaa ttgaaagaaa agcaagaaaa aaccaccaagcattttaaag 240 aagtttggga aaagggcgaa a atg agc aag aaa aat agc gta atttct ggt 291 Met Ser Lys Lys Asn Ser Val Ile Ser Gly 1 5 10 tta atg aatttt ttt agc gaa aag aat gaa cgc tgg ctc tta gcc cac 339 Leu Met Asn PhePhe Ser Glu Lys Asn Glu Arg Trp Leu Leu Ala His 15 20 25 agg cac acg agaggg ttt gtg ata gtg gcg tgg ctt ttt cgg ttt aaa 387 Arg His Thr Arg GlyPhe Val Ile Val Ala Trp Leu Phe Arg Phe Lys 30 35 40 agc att gcg ttt tctatt ttg atc act ctg ttg gtt att tta gtg gat 435 Ser Ile Ala Phe Ser IleLeu Ile Thr Leu Leu Val Ile Leu Val Asp 45 50 55 att tgg gtt tat agc gatgtg cgt cag ttt tta ttg gac act tct agc 483 Ile Trp Val Tyr Ser Asp ValArg Gln Phe Leu Leu Asp Thr Ser Ser 60 65 70 tct ttt att tgg ctt ttg atcgct tta cta atc aag tgg ggc gtg att 531 Ser Phe Ile Trp Leu Leu Ile AlaLeu Leu Ile Lys Trp Gly Val Ile 75 80 85 90 gtc ata agc gca cgt aaa tgctac caa ttc agc caa aaa atg ttt acg 579 Val Ile Ser Ala Arg Lys Cys TyrGln Phe Ser Gln Lys Met Phe Thr 95 100 105 ctc att caa aga aaa agg caaatc aga gag aat tta aaa aac cgc tcc 627 Leu Ile Gln Arg Lys Arg Gln IleArg Glu Asn Leu Lys Asn Arg Ser 110 115 120 aac tac aaa gat acc aaa aatgcg gaa aaa ctc tct agc atc gct gaa 675 Asn Tyr Lys Asp Thr Lys Asn AlaGlu Lys Leu Ser Ser Ile Ala Glu 125 130 135 gaa atc att tca aaa aaa caagaa gag tcc cgc ccc aaa gaa gat tct 723 Glu Ile Ile Ser Lys Lys Gln GluGlu Ser Arg Pro Lys Glu Asp Ser 140 145 150 aat cat gaa aac cat aaa gaaaag ctt tct aac att acc gaa gaa agt 771 Asn His Glu Asn His Lys Glu LysLeu Ser Asn Ile Thr Glu Glu Ser 155 160 165 170 gat tct taaaaaacaagaggaattga aaagctaaaa aggatagggg gggattaccc 827 Asp Ser aaagcatattggaggg 843 <210> SEQ ID NO 4 <211> LENGTH: 172 <212> TYPE: PRT <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 4 Met Ser Lys Lys Asn SerVal Ile Ser Gly Leu Met Asn Phe Phe Ser 1 5 10 15 Glu Lys Asn Glu ArgTrp Leu Leu Ala His Arg His Thr Arg Gly Phe 20 25 30 Val Ile Val Ala TrpLeu Phe Arg Phe Lys Ser Ile Ala Phe Ser Ile 35 40 45 Leu Ile Thr Leu LeuVal Ile Leu Val Asp Ile Trp Val Tyr Ser Asp 50 55 60 Val Arg Gln Phe LeuLeu Asp Thr Ser Ser Ser Phe Ile Trp Leu Leu 65 70 75 80 Ile Ala Leu LeuIle Lys Trp Gly Val Ile Val Ile Ser Ala Arg Lys 85 90 95 Cys Tyr Gln PheSer Gln Lys Met Phe Thr Leu Ile Gln Arg Lys Arg 100 105 110 Gln Ile ArgGlu Asn Leu Lys Asn Arg Ser Asn Tyr Lys Asp Thr Lys 115 120 125 Asn AlaGlu Lys Leu Ser Ser Ile Ala Glu Glu Ile Ile Ser Lys Lys 130 135 140 GlnGlu Glu Ser Arg Pro Lys Glu Asp Ser Asn His Glu Asn His Lys 145 150 155160 Glu Lys Leu Ser Asn Ile Thr Glu Glu Ser Asp Ser 165 170 <210> SEQ IDNO 5 <211> LENGTH: 904 <212> TYPE: DNA <213> ORGANISM: Helicobacterpylori <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (248)...(805)<221> NAME/KEY: sig_peptide <222> LOCATION: (248)...(298) <221>NAME/KEY: mat_peptide <222> LOCATION: (299)...(805) <400> SEQUENCE: 5aaaaaggccc ccatttttaa aagaaaatgg ggggctttaa caaggaagtg aacttgttgt 60atgaagaaaa ttctaaagaa cgcgatcaaa aaagcgcatc aaagtttatc cactatccct 120ctaagttctt cactctatgc tataatctct gttttaaaac attatggcgt gttagaagat 180attcagcaaa acccttccaa accaaccaat ctaaagaaag aaaccattca aggaacgcat 240tgatttg atg aat aaa cca ttt tta atc tta ctc ata gcc cta att gtc 289 MetAsn Lys Pro Phe Leu Ile Leu Leu Ile Ala Leu Ile Val -15 -10 -5 ttt agcggc tgt aac atg aga aaa tat ttc aaa ccc gct aaa cac caa 337 Phe Ser GlyCys Asn Met Arg Lys Tyr Phe Lys Pro Ala Lys His Gln 1 5 10 gtt aaa ggcgaa gcg tat ttc cct aat cat ttg caa gaa agt atc gtt 385 Val Lys Gly GluAla Tyr Phe Pro Asn His Leu Gln Glu Ser Ile Val 15 20 25 tcg tct aat cgttat gga gcc att ttg aaa aat gga gcg gtt ata ggc 433 Ser Ser Asn Arg TyrGly Ala Ile Leu Lys Asn Gly Ala Val Ile Gly 30 35 40 45 gat aaa ggt ttaacg cag cta aga atc ggt aag aat ttc aat tat gaa 481 Asp Lys Gly Leu ThrGln Leu Arg Ile Gly Lys Asn Phe Asn Tyr Glu 50 55 60 agc agt ttt tta aatgag agt cag ggg ttt ttc atc ctt gcg caa gat 529 Ser Ser Phe Leu Asn GluSer Gln Gly Phe Phe Ile Leu Ala Gln Asp 65 70 75 tgt ttg aac aag att gataaa aaa aca agc aaa aac aag gtg gct aaa 577 Cys Leu Asn Lys Ile Asp LysLys Thr Ser Lys Asn Lys Val Ala Lys 80 85 90 agt gag gaa acg gag ctg aaatta aag ggc gtt gaa gcc gaa gtc caa 625 Ser Glu Glu Thr Glu Leu Lys LeuLys Gly Val Glu Ala Glu Val Gln 95 100 105 gat aaa gtc tgt cat caa gtggaa ttg att agc aat aac cct aac gcc 673 Asp Lys Val Cys His Gln Val GluLeu Ile Ser Asn Asn Pro Asn Ala 110 115 120 125 agc caa caa tct atc gttatc cct ttg gag act ttt gcc ttg agc gca 721 Ser Gln Gln Ser Ile Val IlePro Leu Glu Thr Phe Ala Leu Ser Ala 130 135 140 agc gtt aaa ggg aat ctttta gcg gtg gtg ttt agc gga caa ttc agc 769 Ser Val Lys Gly Asn Leu LeuAla Val Val Phe Ser Gly Gln Phe Ser 145 150 155 gaa ttt ata cga cat cacttc tca aaa att gct ttt tagtgagaaa 815 Glu Phe Ile Arg His His Phe SerLys Ile Ala Phe 160 165 ggttccccaa gcaccacgat caattcttta atggcgaatgcctattttta atggatacgg 875 tccttgtgtt tcccccaagc caaaatggg 904 <210> SEQID NO 6 <211> LENGTH: 186 <212> TYPE: PRT <213> ORGANISM: Helicobacterpylori <400> SEQUENCE: 6 Met Asn Lys Pro Phe Leu Ile Leu Leu Ile Ala LeuIle Val Phe Ser 1 5 10 15 Gly Cys Asn Met Arg Lys Tyr Phe Lys Pro AlaLys His Gln Val Lys 20 25 30 Gly Glu Ala Tyr Phe Pro Asn His Leu Gln GluSer Ile Val Ser Ser 35 40 45 Asn Arg Tyr Gly Ala Ile Leu Lys Asn Gly AlaVal Ile Gly Asp Lys 50 55 60 Gly Leu Thr Gln Leu Arg Ile Gly Lys Asn PheAsn Tyr Glu Ser Ser 65 70 75 80 Phe Leu Asn Glu Ser Gln Gly Phe Phe IleLeu Ala Gln Asp Cys Leu 85 90 95 Asn Lys Ile Asp Lys Lys Thr Ser Lys AsnLys Val Ala Lys Ser Glu 100 105 110 Glu Thr Glu Leu Lys Leu Lys Gly ValGlu Ala Glu Val Gln Asp Lys 115 120 125 Val Cys His Gln Val Glu Leu IleSer Asn Asn Pro Asn Ala Ser Gln 130 135 140 Gln Ser Ile Val Ile Pro LeuGlu Thr Phe Ala Leu Ser Ala Ser Val 145 150 155 160 Lys Gly Asn Leu LeuAla Val Val Phe Ser Gly Gln Phe Ser Glu Phe 165 170 175 Ile Arg His HisPhe Ser Lys Ile Ala Phe 180 185 <210> SEQ ID NO 7 <211> LENGTH: 874<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: CDS <222> LOCATION: (146)...(802) <221> NAME/KEY: sig_peptide<222> LOCATION: (146)...(208) <221> NAME/KEY: mat_peptide <222>LOCATION: (209)...(802) <400> SEQUENCE: 7 gcaaaaattt ggctggaaagcccctaggtt ggatccaatt ttgataagga tttttaaccg 60 gcaatttaaa aactattactccctgggatg gttcataatg caaaaaagcc aacgcaactt 120 aatacttcat taaggtttaatcaca atg gat aaa aac aac aac acg aat ctt 172 Met Asp Lys Asn Asn AsnThr Asn Leu -20 -15 att tta gcg atc gct ctg tct ttc ttg ttt atc gct ctttat agg tat 220 Ile Leu Ala Ile Ala Leu Ser Phe Leu Phe Ile Ala Leu TyrArg Tyr -10 -5 1 ttt ttc caa aaa cca aac aaa aca aca acc caa acc aca aagcaa gaa 268 Phe Phe Gln Lys Pro Asn Lys Thr Thr Thr Gln Thr Thr Lys GlnGlu 5 10 15 20 aca gcc aac aac cac aca gca aca agt cct aac gcg ccc aacgcc caa 316 Thr Ala Asn Asn His Thr Ala Thr Ser Pro Asn Ala Pro Asn AlaGln 25 30 35 aat ttt agc gtt act caa acc atc ccc caa gag agt ttg tta agcacg 364 Asn Phe Ser Val Thr Gln Thr Ile Pro Gln Glu Ser Leu Leu Ser Thr40 45 50 att tct ttt gag cat gcc agg att gaa att gat tct tta ggg cgc atc412 Ile Ser Phe Glu His Ala Arg Ile Glu Ile Asp Ser Leu Gly Arg Ile 5560 65 aaa cag gtt tat ctc aag gat aaa aag tat cta acc cct aaa caa aag460 Lys Gln Val Tyr Leu Lys Asp Lys Lys Tyr Leu Thr Pro Lys Gln Lys 7075 80 ggc ttt tta gag cat gtg agc cat ctt ttt aac ccc aaa gct aac ccg508 Gly Phe Leu Glu His Val Ser His Leu Phe Asn Pro Lys Ala Asn Pro 8590 95 100 caa ccc ccc cta aaa gag ctc ccc ctt tta gcg gcc gat aaa ctcaag 556 Gln Pro Pro Leu Lys Glu Leu Pro Leu Leu Ala Ala Asp Lys Leu Lys105 110 115 cct tta gaa gtg cgt ttt tta gac ccc acg ctc aat aac aaa gcgttc 604 Pro Leu Glu Val Arg Phe Leu Asp Pro Thr Leu Asn Asn Lys Ala Phe120 125 130 aac acc cct tat agt gct tca aaa acc act ctt ggg cct aat gaacag 652 Asn Thr Pro Tyr Ser Ala Ser Lys Thr Thr Leu Gly Pro Asn Glu Gln135 140 145 ctt gtt tta acc caa gat tta ggc gct ctt acc atc att aaa accctg 700 Leu Val Leu Thr Gln Asp Leu Gly Ala Leu Thr Ile Ile Lys Thr Leu150 155 160 act ttt tat gat gat ttg cat tat gat tta aga atc gcc ttc aaatcg 748 Thr Phe Tyr Asp Asp Leu His Tyr Asp Leu Arg Ile Ala Phe Lys Ser165 170 175 180 cct aac aat att atc cct agc tat gtg atc act aat ggt tacaga ccg 796 Pro Asn Asn Ile Ile Pro Ser Tyr Val Ile Thr Asn Gly Tyr ArgPro 185 190 195 ggt ggc tgatttggac agctacacct tttcgggcgt gcatattagaaaacaacgac 852 Gly Gly gaaaaattaa gggtattgaa ca 874 <210> SEQ ID NO 8<211> LENGTH: 219 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 8 Met Asp Lys Asn Asn Asn Thr Asn Leu Ile Leu Ala IleAla Leu Ser 1 5 10 15 Phe Leu Phe Ile Ala Leu Tyr Arg Tyr Phe Phe GlnLys Pro Asn Lys 20 25 30 Thr Thr Thr Gln Thr Thr Lys Gln Glu Thr Ala AsnAsn His Thr Ala 35 40 45 Thr Ser Pro Asn Ala Pro Asn Ala Gln Asn Phe SerVal Thr Gln Thr 50 55 60 Ile Pro Gln Glu Ser Leu Leu Ser Thr Ile Ser PheGlu His Ala Arg 65 70 75 80 Ile Glu Ile Asp Ser Leu Gly Arg Ile Lys GlnVal Tyr Leu Lys Asp 85 90 95 Lys Lys Tyr Leu Thr Pro Lys Gln Lys Gly PheLeu Glu His Val Ser 100 105 110 His Leu Phe Asn Pro Lys Ala Asn Pro GlnPro Pro Leu Lys Glu Leu 115 120 125 Pro Leu Leu Ala Ala Asp Lys Leu LysPro Leu Glu Val Arg Phe Leu 130 135 140 Asp Pro Thr Leu Asn Asn Lys AlaPhe Asn Thr Pro Tyr Ser Ala Ser 145 150 155 160 Lys Thr Thr Leu Gly ProAsn Glu Gln Leu Val Leu Thr Gln Asp Leu 165 170 175 Gly Ala Leu Thr IleIle Lys Thr Leu Thr Phe Tyr Asp Asp Leu His 180 185 190 Tyr Asp Leu ArgIle Ala Phe Lys Ser Pro Asn Asn Ile Ile Pro Ser 195 200 205 Tyr Val IleThr Asn Gly Tyr Arg Pro Gly Gly 210 215 <210> SEQ ID NO 9 <211> LENGTH:761 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE:<221> NAME/KEY: CDS <222> LOCATION: (303)...(758) <221> NAME/KEY:sig_peptide <222> LOCATION: (303)...(362) <221> NAME/KEY: mat_peptide<222> LOCATION: (363)...(758) <400> SEQUENCE: 9 taaaaaatgc aaaaacatttttgggttatt aaaaagattc cttctaggag tcgaacctgc 60 caggcatgca aggtagctttcgcgagcttt gagatccctt caaacgcttt gattagaaac 120 gggaaagatt acctggtgtttgtgagaacg cctaaaggtt ttaggcctgt ggtggttcaa 180 gttttagaag agcgcagcaagatttttatc gtgaacgctc aaaatttaca ccctaatgac 240 agcgtggcag tggggtcattgatagggtta aaaggcatga tcaacaattt aggggaagaa 300 ta atg ctc gct tcc attatt gaa ttt tcc tta cgc cag cga ata atc 347 Met Leu Ala Ser Ile Ile GluPhe Ser Leu Arg Gln Arg Ile Ile -20 -15 -10 gtg att gtt ggc gcg att cttatt ttg ttt ttt ggg act tat agt ttt 395 Val Ile Val Gly Ala Ile Leu IleLeu Phe Phe Gly Thr Tyr Ser Phe -5 1 5 10 atc cac act cca gta gat gctttc ccg gat att tcg ccc act caa gtc 443 Ile His Thr Pro Val Asp Ala PhePro Asp Ile Ser Pro Thr Gln Val 15 20 25 aaa atc att tta aaa ctc ccc ggttct agc cct gaa gaa atg gaa aat 491 Lys Ile Ile Leu Lys Leu Pro Gly SerSer Pro Glu Glu Met Glu Asn 30 35 40 aac atc gtg cgc cct tta gaa ttg gagctt tta ggc ttg aaa ggg caa 539 Asn Ile Val Arg Pro Leu Glu Leu Glu LeuLeu Gly Leu Lys Gly Gln 45 50 55 aaa tct tta aga agt att tca aaa tat tctatt tca gac atc acg ata 587 Lys Ser Leu Arg Ser Ile Ser Lys Tyr Ser IleSer Asp Ile Thr Ile 60 65 70 75 gat ttt gat gac agc gtg gat att tat ttagcg aga aac att gtt aat 635 Asp Phe Asp Asp Ser Val Asp Ile Tyr Leu AlaArg Asn Ile Val Asn 80 85 90 gag cgc ttg agc agc gtg atg aaa gat tta cccgtg ggg gtt gaa agg 683 Glu Arg Leu Ser Ser Val Met Lys Asp Leu Pro ValGly Val Glu Arg 95 100 105 ggc atg gcg ccc att gtt acg ccg cta tca aatatc ttt atg ttt cac 731 Gly Met Ala Pro Ile Val Thr Pro Leu Ser Asn IlePhe Met Phe His 110 115 120 tat tgg atg ggc cat atc cct gga gaa tag 761Tyr Trp Met Gly His Ile Pro Gly Glu 125 130 <210> SEQ ID NO 10 <211>LENGTH: 152 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 10 Met Leu Ala Ser Ile Ile Glu Phe Ser Leu Arg Gln Arg Ile IleVal 1 5 10 15 Ile Val Gly Ala Ile Leu Ile Leu Phe Phe Gly Thr Tyr SerPhe Ile 20 25 30 His Thr Pro Val Asp Ala Phe Pro Asp Ile Ser Pro Thr GlnVal Lys 35 40 45 Ile Ile Leu Lys Leu Pro Gly Ser Ser Pro Glu Glu Met GluAsn Asn 50 55 60 Ile Val Arg Pro Leu Glu Leu Glu Leu Leu Gly Leu Lys GlyGln Lys 65 70 75 80 Ser Leu Arg Ser Ile Ser Lys Tyr Ser Ile Ser Asp IleThr Ile Asp 85 90 95 Phe Asp Asp Ser Val Asp Ile Tyr Leu Ala Arg Asn IleVal Asn Glu 100 105 110 Arg Leu Ser Ser Val Met Lys Asp Leu Pro Val GlyVal Glu Arg Gly 115 120 125 Met Ala Pro Ile Val Thr Pro Leu Ser Asn IlePhe Met Phe His Tyr 130 135 140 Trp Met Gly His Ile Pro Gly Glu 145 150<210> SEQ ID NO 11 <211> LENGTH: 392 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION:(25)...(366) <400> SEQUENCE: 11 gaattgaacc atgctgaagg gtgt atg gta ttagga agc tta tac cat cat 51 Met Val Leu Gly Ser Leu Tyr His His 1 5 ggctta ggc acg cct aag gat tca aga aag gct ctt gat ttg tat gaa 99 Gly LeuGly Thr Pro Lys Asp Ser Arg Lys Ala Leu Asp Leu Tyr Glu 10 15 20 25 aaagct tgc gat tta aaa gac agc ccc ggg tgc att aat gca gga tac 147 Lys AlaCys Asp Leu Lys Asp Ser Pro Gly Cys Ile Asn Ala Gly Tyr 30 35 40 atg tatggt gta gca aag aat ttt aag gag gct att gtt cgt tat tct 195 Met Tyr GlyVal Ala Lys Asn Phe Lys Glu Ala Ile Val Arg Tyr Ser 45 50 55 aag gca tgcgaa ttg aaa gat ggc agg ggg tgt tat aat tta ggg gtt 243 Lys Ala Cys GluLeu Lys Asp Gly Arg Gly Cys Tyr Asn Leu Gly Val 60 65 70 atg caa tac aacgcc caa ggc aca gca aag gac gaa aag caa gcg gta 291 Met Gln Tyr Asn AlaGln Gly Thr Ala Lys Asp Glu Lys Gln Ala Val 75 80 85 gaa aac ttt aaa aaaggt tgc aaa tca agc gtt aaa gaa gca tgc gac 339 Glu Asn Phe Lys Lys GlyCys Lys Ser Ser Val Lys Glu Ala Cys Asp 90 95 100 105 gct ctc aag gaatta aaa ata gaa ctt tagttttaac gaagttaagc 386 Ala Leu Lys Glu Leu LysIle Glu Leu 110 taaagg 392 <210> SEQ ID NO 12 <211> LENGTH: 114 <212>TYPE: PRT <213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 12 Met ValLeu Gly Ser Leu Tyr His His Gly Leu Gly Thr Pro Lys Asp 1 5 10 15 SerArg Lys Ala Leu Asp Leu Tyr Glu Lys Ala Cys Asp Leu Lys Asp 20 25 30 SerPro Gly Cys Ile Asn Ala Gly Tyr Met Tyr Gly Val Ala Lys Asn 35 40 45 PheLys Glu Ala Ile Val Arg Tyr Ser Lys Ala Cys Glu Leu Lys Asp 50 55 60 GlyArg Gly Cys Tyr Asn Leu Gly Val Met Gln Tyr Asn Ala Gln Gly 65 70 75 80Thr Ala Lys Asp Glu Lys Gln Ala Val Glu Asn Phe Lys Lys Gly Cys 85 90 95Lys Ser Ser Val Lys Glu Ala Cys Asp Ala Leu Lys Glu Leu Lys Ile 100 105110 Glu Leu <210> SEQ ID NO 13 <211> LENGTH: 982 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY: CDS <222>LOCATION: (117)...(911) <221> NAME/KEY: sig_peptide <222> LOCATION:(117)...(167) <221> NAME/KEY: mat_peptide <222> LOCATION: (168)...(911)<400> SEQUENCE: 13 ccacatttaa ggtagaaacc actcaattag atgtaaaaattccaaacggc aaccaaaaaa 60 tggttaaaaa ggacacaata aaccccaaaa atgaaatttaaatatatggg aactta atg 119 Met aga att ttt ttt gtt atc atg gga ctt gtgttt ttt ggt tgc acc agt 167 Arg Ile Phe Phe Val Ile Met Gly Leu Val PhePhe Gly Cys Thr Ser -15 -10 -5 aag gtg cat gag atg aaa aaa agc cct tgcacc ttg tat gaa aac agg 215 Lys Val His Glu Met Lys Lys Ser Pro Cys ThrLeu Tyr Glu Asn Arg 1 5 10 15 tta aat ctc gca gaa atc ttt cac aag cgagca att gat cta ttt aga 263 Leu Asn Leu Ala Glu Ile Phe His Lys Arg AlaIle Asp Leu Phe Arg 20 25 30 gag ctt tta agc cac caa gaa aag cat tta gaaaac aag ctt tct ggt 311 Glu Leu Leu Ser His Gln Glu Lys His Leu Glu AsnLys Leu Ser Gly 35 40 45 ttt tcg gtg agt gat ttg gac atg caa agc gtg tttcgg ctg gaa aga 359 Phe Ser Val Ser Asp Leu Asp Met Gln Ser Val Phe ArgLeu Glu Arg 50 55 60 aac cgc ttg aaa atc gct tac aag ctc tta ggc ttg atgagt ttt atc 407 Asn Arg Leu Lys Ile Ala Tyr Lys Leu Leu Gly Leu Met SerPhe Ile 65 70 75 80 gct ctt att tta gcg atc gtg tta atc agt ctt cta ccctta caa aaa 455 Ala Leu Ile Leu Ala Ile Val Leu Ile Ser Leu Leu Pro LeuGln Lys 85 90 95 acc gaa cac cat ttc gtg gat ttt tta aac cag gac aag cattac gtc 503 Thr Glu His His Phe Val Asp Phe Leu Asn Gln Asp Lys His TyrVal 100 105 110 att atc caa aga gcg gat aaa agc att tcc agt aat gaa gcgttg gct 551 Ile Ile Gln Arg Ala Asp Lys Ser Ile Ser Ser Asn Glu Ala LeuAla 115 120 125 cgt tcg ctc att ggg gcg tat gtg tta aac cga gag agc attaac cgc 599 Arg Ser Leu Ile Gly Ala Tyr Val Leu Asn Arg Glu Ser Ile AsnArg 130 135 140 att gac gat aaa tcg cgc tat gaa ttg gtg cgc ttg caa agcagt tct 647 Ile Asp Asp Lys Ser Arg Tyr Glu Leu Val Arg Leu Gln Ser SerSer 145 150 155 160 aaa gtg tgg caa cgc ttt gaa gat ttg att aaa acc caaaac agc att 695 Lys Val Trp Gln Arg Phe Glu Asp Leu Ile Lys Thr Gln AsnSer Ile 165 170 175 tat gtg caa agc cat ttg gaa aga gaa gtc cat atc gtcaat att gcg 743 Tyr Val Gln Ser His Leu Glu Arg Glu Val His Ile Val AsnIle Ala 180 185 190 atc tat cag caa gac aat aac ccc att gcg agc gtc tccatt gcc gct 791 Ile Tyr Gln Gln Asp Asn Asn Pro Ile Ala Ser Val Ser IleAla Ala 195 200 205 aaa ctt ttg aat gaa aac aag ctg gtg tat gaa aag cgttat aaa atc 839 Lys Leu Leu Asn Glu Asn Lys Leu Val Tyr Glu Lys Arg TyrLys Ile 210 215 220 gta ttg agt tat ttg ttt gac acc ccg atg aat tca agcttg caa gct 887 Val Leu Ser Tyr Leu Phe Asp Thr Pro Met Asn Ser Ser LeuGln Ala 225 230 235 240 tgc aag ctc tca ggc ttc ata gtt tgacatgacatatagatgag ctttatgcgg 941 Cys Lys Leu Ser Gly Phe Ile Val 245 tacgattatcacagaatggc taacgcagca ggcaccgagt a 982 <210> SEQ ID NO 14 <211> LENGTH:265 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori <400> SEQUENCE:14 Met Arg Ile Phe Phe Val Ile Met Gly Leu Val Phe Phe Gly Cys Thr 1 510 15 Ser Lys Val His Glu Met Lys Lys Ser Pro Cys Thr Leu Tyr Glu Asn 2025 30 Arg Leu Asn Leu Ala Glu Ile Phe His Lys Arg Ala Ile Asp Leu Phe 3540 45 Arg Glu Leu Leu Ser His Gln Glu Lys His Leu Glu Asn Lys Leu Ser 5055 60 Gly Phe Ser Val Ser Asp Leu Asp Met Gln Ser Val Phe Arg Leu Glu 6570 75 80 Arg Asn Arg Leu Lys Ile Ala Tyr Lys Leu Leu Gly Leu Met Ser Phe85 90 95 Ile Ala Leu Ile Leu Ala Ile Val Leu Ile Ser Leu Leu Pro Leu Gln100 105 110 Lys Thr Glu His His Phe Val Asp Phe Leu Asn Gln Asp Lys HisTyr 115 120 125 Val Ile Ile Gln Arg Ala Asp Lys Ser Ile Ser Ser Asn GluAla Leu 130 135 140 Ala Arg Ser Leu Ile Gly Ala Tyr Val Leu Asn Arg GluSer Ile Asn 145 150 155 160 Arg Ile Asp Asp Lys Ser Arg Tyr Glu Leu ValArg Leu Gln Ser Ser 165 170 175 Ser Lys Val Trp Gln Arg Phe Glu Asp LeuIle Lys Thr Gln Asn Ser 180 185 190 Ile Tyr Val Gln Ser His Leu Glu ArgGlu Val His Ile Val Asn Ile 195 200 205 Ala Ile Tyr Gln Gln Asp Asn AsnPro Ile Ala Ser Val Ser Ile Ala 210 215 220 Ala Lys Leu Leu Asn Glu AsnLys Leu Val Tyr Glu Lys Arg Tyr Lys 225 230 235 240 Ile Val Leu Ser TyrLeu Phe Asp Thr Pro Met Asn Ser Ser Leu Gln 245 250 255 Ala Cys Lys LeuSer Gly Phe Ile Val 260 265 <210> SEQ ID NO 15 <211> LENGTH: 760 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: CDS <222> LOCATION: (236)...(577) <221> NAME/KEY: sig_peptide<222> LOCATION: (236)...(355) <221> NAME/KEY: mat_peptide <222>LOCATION: (356)...(577) <400> SEQUENCE: 15 ctaaaaatgg aagcctaaaaaggggtaaaa aagctttaaa aagggggcaa aaaattgaag 60 cgatttcaaa aaaaaaaaaaaaaaacaatt tcagtttctt attagctaga tttgattaga 120 ataaaaagct tttatgtgtttaaacttcat tgtcttaaaa cttttaagag caattttaaa 180 attcgttggc gtataatatcaaatttgaat gaactgacta aaagggtttt aaata atg 238 Met -40 gct gaa aat tctttc aaa aat gtt tcc aca caa ccc aaa cca ttt ttc 286 Ala Glu Asn Ser PheLys Asn Val Ser Thr Gln Pro Lys Pro Phe Phe -35 -30 -25 tta tta cca gttaaa acc ctg ttt ctt tta gga ggc gtt ttt agc gcg 334 Leu Leu Pro Val LysThr Leu Phe Leu Leu Gly Gly Val Phe Ser Ala -20 -15 -10 ttt ttt atc cttgtt gct ggc ttg gtt ttt ttt aat tac act aat tca 382 Phe Phe Ile Leu ValAla Gly Leu Val Phe Phe Asn Tyr Thr Asn Ser -5 1 5 atg gac cat gcg attttt aac ttg atg cgt tca aac tct tct aac cct 430 Met Asp His Ala Ile PheAsn Leu Met Arg Ser Asn Ser Ser Asn Pro 10 15 20 25 att tta gat caa acgctc cga cgc gtt gtt ttt tta ggc tct tct caa 478 Ile Leu Asp Gln Thr LeuArg Arg Val Val Phe Leu Gly Ser Ser Gln 30 35 40 ttc gtg ttg cct ttg agcttg tta gtg ggg gtt ttt tta agc ttg tat 526 Phe Val Leu Pro Leu Ser LeuLeu Val Gly Val Phe Leu Ser Leu Tyr 45 50 55 cgt aaa aat tta gca ctt ggggtg tgg ttt gtg cta aag cgt gat ctt 574 Arg Lys Asn Leu Ala Leu Gly ValTrp Phe Val Leu Lys Arg Asp Leu 60 65 70 att tgaagccctt ttagaatctttaaaacacct tttggcacac tccattcaat 627 Ile gggttttcgg cacaggctaatttccctagc actatcgggc tttctttgac gggaatttta 687 tggggtgctt ggttttaattaataccccat ttgttccacg gatcaaaaat ttcaaaaaca 747 tttttttcgt aaa 760 <210>SEQ ID NO 16 <211> LENGTH: 114 <212> TYPE: PRT <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 16 Met Ala Glu Asn Ser Phe Lys AsnVal Ser Thr Gln Pro Lys Pro Phe 1 5 10 15 Phe Leu Leu Pro Val Lys ThrLeu Phe Leu Leu Gly Gly Val Phe Ser 20 25 30 Ala Phe Phe Ile Leu Val AlaGly Leu Val Phe Phe Asn Tyr Thr Asn 35 40 45 Ser Met Asp His Ala Ile PheAsn Leu Met Arg Ser Asn Ser Ser Asn 50 55 60 Pro Ile Leu Asp Gln Thr LeuArg Arg Val Val Phe Leu Gly Ser Ser 65 70 75 80 Gln Phe Val Leu Pro LeuSer Leu Leu Val Gly Val Phe Leu Ser Leu 85 90 95 Tyr Arg Lys Asn Leu AlaLeu Gly Val Trp Phe Val Leu Lys Arg Asp 100 105 110 Leu Ile <210> SEQ IDNO 17 <211> LENGTH: 939 <212> TYPE: DNA <213> ORGANISM: Helicobacterpylori <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (37)...(816)<221> NAME/KEY: sig_peptide <222> LOCATION: (37)...(138) <221> NAME/KEY:mat_peptide <222> LOCATION: (139)...(816) <400> SEQUENCE: 17 agtttgcaaccctggatgag agtagtggca gagttt atg ctg att ccg tta aaa 54 Met Leu Ile ProLeu Lys -30 aca ttc cta aaa ata tta ttg aaa ata ttc cta aaa act ttc caaaag 102 Thr Phe Leu Lys Ile Leu Leu Lys Ile Phe Leu Lys Thr Phe Gln Lys-25 -20 -15 att tgg ata gtt tgc gtt gtt att tgg ggg ttg ggt tgt agt ttttta 150 Ile Trp Ile Val Cys Val Val Ile Trp Gly Leu Gly Cys Ser Phe Leu-10 -5 1 aac gct aac agc att caa tta gaa gaa acg ctc aga cga aat cct aaa198 Asn Ala Asn Ser Ile Gln Leu Glu Glu Thr Leu Arg Arg Asn Pro Lys 5 1015 20 aat ctt att tgg caa cac ttt aaa aag aag ttt aaa aag agc aac acg246 Asn Leu Ile Trp Gln His Phe Lys Lys Lys Phe Lys Lys Ser Asn Thr 2530 35 atc cct tat gcc cca aac agc cgt tgg aaa tat tta ggc acg agc ata294 Ile Pro Tyr Ala Pro Asn Ser Arg Trp Lys Tyr Leu Gly Thr Ser Ile 4045 50 ggg att tta ggc gtg tcg ttg gtg ata ggg att gtg ggg ttg tat ctc342 Gly Ile Leu Gly Val Ser Leu Val Ile Gly Ile Val Gly Leu Tyr Leu 5560 65 atg cca gag agc gta acg aat tgg gat aga gaa aag ttt ggc gtc aaa390 Met Pro Glu Ser Val Thr Asn Trp Asp Arg Glu Lys Phe Gly Val Lys 7075 80 agt tgg ttt gaa aat gtc cgc atg ggg cca aaa ctg gac aat gat agt438 Ser Trp Phe Glu Asn Val Arg Met Gly Pro Lys Leu Asp Asn Asp Ser 8590 95 100 ttt att ttc aat gaa att ttg cac cct tat ttt ggg gct atg tattat 486 Phe Ile Phe Asn Glu Ile Leu His Pro Tyr Phe Gly Ala Met Tyr Tyr105 110 115 atg caa ccg cgc atg gct ggg ttt ggc tgg atg gca tca gcg tttttt 534 Met Gln Pro Arg Met Ala Gly Phe Gly Trp Met Ala Ser Ala Phe Phe120 125 130 tct ttt atc act tct acg ctt ttt tgg gaa tat ggc ttg gaa gcgttt 582 Ser Phe Ile Thr Ser Thr Leu Phe Trp Glu Tyr Gly Leu Glu Ala Phe135 140 145 gtg gaa gtg cct agc tgg cag gat tta gtg atc acg cct tta ttaggc 630 Val Glu Val Pro Ser Trp Gln Asp Leu Val Ile Thr Pro Leu Leu Gly150 155 160 tcc att tta ggg gaa ggg ttt tat cag ctc acg cgc tat atc caacgg 678 Ser Ile Leu Gly Glu Gly Phe Tyr Gln Leu Thr Arg Tyr Ile Gln Arg165 170 175 180 aac caa ggc aag ctt ttt ggc tct tta ttt tta ggg cgt ttagcc atc 726 Asn Gln Gly Lys Leu Phe Gly Ser Leu Phe Leu Gly Arg Leu AlaIle 185 190 195 gct ctt atg gat cct atc ggt ttt gtg atc agg gat tta gggctt ggg 774 Ala Leu Met Asp Pro Ile Gly Phe Val Ile Arg Asp Leu Gly LeuGly 200 205 210 gaa gct tta gga ttc ata ata aac atg aaa tcc gtt cca gtt816 Glu Ala Leu Gly Phe Ile Ile Asn Met Lys Ser Val Pro Val 215 220 225taagccctaa tggcttgaat ttgacttaca aattctaaag aataacccgc atcaagcgac 876tttaaaaaca ttttcaccaa tgaaataccc cgctctatct atgaaccgac tatacaaacg 936aac 939 <210> SEQ ID NO 18 <211> LENGTH: 260 <212> TYPE: PRT <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 18 Met Leu Ile Pro Leu LysThr Phe Leu Lys Ile Leu Leu Lys Ile Phe 1 5 10 15 Leu Lys Thr Phe GlnLys Ile Trp Ile Val Cys Val Val Ile Trp Gly 20 25 30 Leu Gly Cys Ser PheLeu Asn Ala Asn Ser Ile Gln Leu Glu Glu Thr 35 40 45 Leu Arg Arg Asn ProLys Asn Leu Ile Trp Gln His Phe Lys Lys Lys 50 55 60 Phe Lys Lys Ser AsnThr Ile Pro Tyr Ala Pro Asn Ser Arg Trp Lys 65 70 75 80 Tyr Leu Gly ThrSer Ile Gly Ile Leu Gly Val Ser Leu Val Ile Gly 85 90 95 Ile Val Gly LeuTyr Leu Met Pro Glu Ser Val Thr Asn Trp Asp Arg 100 105 110 Glu Lys PheGly Val Lys Ser Trp Phe Glu Asn Val Arg Met Gly Pro 115 120 125 Lys LeuAsp Asn Asp Ser Phe Ile Phe Asn Glu Ile Leu His Pro Tyr 130 135 140 PheGly Ala Met Tyr Tyr Met Gln Pro Arg Met Ala Gly Phe Gly Trp 145 150 155160 Met Ala Ser Ala Phe Phe Ser Phe Ile Thr Ser Thr Leu Phe Trp Glu 165170 175 Tyr Gly Leu Glu Ala Phe Val Glu Val Pro Ser Trp Gln Asp Leu Val180 185 190 Ile Thr Pro Leu Leu Gly Ser Ile Leu Gly Glu Gly Phe Tyr GlnLeu 195 200 205 Thr Arg Tyr Ile Gln Arg Asn Gln Gly Lys Leu Phe Gly SerLeu Phe 210 215 220 Leu Gly Arg Leu Ala Ile Ala Leu Met Asp Pro Ile GlyPhe Val Ile 225 230 235 240 Arg Asp Leu Gly Leu Gly Glu Ala Leu Gly PheIle Ile Asn Met Lys 245 250 255 Ser Val Pro Val 260 <210> SEQ ID NO 19<211> LENGTH: 603 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (1)...(600) <221>NAME/KEY: sig_peptide <222> LOCATION: (1)...(63) <221> NAME/KEY:mat_peptide <222> LOCATION: (64)...(600) <400> SEQUENCE: 19 atg aaa agattt gat ttg ttt tta tca ctc atg ggt gtt tgc gtt tgc 48 Met Lys Arg PheAsp Leu Phe Leu Ser Leu Met Gly Val Cys Val Cys -20 -15 -10 gtt caa acttac gcc gag caa gat tac ttt ttt agg gat ttt aaa tct 96 Val Gln Thr TyrAla Glu Gln Asp Tyr Phe Phe Arg Asp Phe Lys Ser -5 1 5 10 aaa gac ttgccc caa aaa ctc cat ctt gat aaa aag ctc tcc caa aca 144 Lys Asp Leu ProGln Lys Leu His Leu Asp Lys Lys Leu Ser Gln Thr 15 20 25 ata cag cca tgcgcg caa ctt aac gca tca aaa cac tac act gct acc 192 Ile Gln Pro Cys AlaGln Leu Asn Ala Ser Lys His Tyr Thr Ala Thr 30 35 40 ggg gtt aga gag cctgat aaa tgc aca aag agt ttt aaa aaa tcc gct 240 Gly Val Arg Glu Pro AspLys Cys Thr Lys Ser Phe Lys Lys Ser Ala 45 50 55 atc atg tcc tat gac ttagcg cta ggc tat tgg gtg agc caa aac aaa 288 Ile Met Ser Tyr Asp Leu AlaLeu Gly Tyr Trp Val Ser Gln Asn Lys 60 65 70 75 caa tac ggc ttt aaa gctata gaa aat tta aac gct tgg gct aaa gag 336 Gln Tyr Gly Phe Lys Ala IleGlu Asn Leu Asn Ala Trp Ala Lys Glu 80 85 90 ctt caa agc gtg gat act tatcag agc gag gat aat atc aat ttc tac 384 Leu Gln Ser Val Asp Thr Tyr GlnSer Glu Asp Asn Ile Asn Phe Tyr 95 100 105 atg cct tat atg aac atg gcttat tgg ttt gtc aaa aag gca ttc cct 432 Met Pro Tyr Met Asn Met Ala TyrTrp Phe Val Lys Lys Ala Phe Pro 110 115 120 agc cca gaa tac gaa gat ttcgtt aag cgg atg cgc caa tat tct caa 480 Ser Pro Glu Tyr Glu Asp Phe ValLys Arg Met Arg Gln Tyr Ser Gln 125 130 135 tca gct ctt aac act aac catggg gcg tgg ggc att ctc ttt gat gtg 528 Ser Ala Leu Asn Thr Asn His GlyAla Trp Gly Ile Leu Phe Asp Val 140 145 150 155 agc tct gca cta gcg ttagat gat cat gct ctt ttg cac aat agc gct 576 Ser Ser Ala Leu Ala Leu AspAsp His Ala Leu Leu His Asn Ser Ala 160 165 170 aat cgg tgg cag gat tgggat att taa 603 Asn Arg Trp Gln Asp Trp Asp Ile 175 <210> SEQ ID NO 20<211> LENGTH: 200 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 20 Met Lys Arg Phe Asp Leu Phe Leu Ser Leu Met Gly ValCys Val Cys 1 5 10 15 Val Gln Thr Tyr Ala Glu Gln Asp Tyr Phe Phe ArgAsp Phe Lys Ser 20 25 30 Lys Asp Leu Pro Gln Lys Leu His Leu Asp Lys LysLeu Ser Gln Thr 35 40 45 Ile Gln Pro Cys Ala Gln Leu Asn Ala Ser Lys HisTyr Thr Ala Thr 50 55 60 Gly Val Arg Glu Pro Asp Lys Cys Thr Lys Ser PheLys Lys Ser Ala 65 70 75 80 Ile Met Ser Tyr Asp Leu Ala Leu Gly Tyr TrpVal Ser Gln Asn Lys 85 90 95 Gln Tyr Gly Phe Lys Ala Ile Glu Asn Leu AsnAla Trp Ala Lys Glu 100 105 110 Leu Gln Ser Val Asp Thr Tyr Gln Ser GluAsp Asn Ile Asn Phe Tyr 115 120 125 Met Pro Tyr Met Asn Met Ala Tyr TrpPhe Val Lys Lys Ala Phe Pro 130 135 140 Ser Pro Glu Tyr Glu Asp Phe ValLys Arg Met Arg Gln Tyr Ser Gln 145 150 155 160 Ser Ala Leu Asn Thr AsnHis Gly Ala Trp Gly Ile Leu Phe Asp Val 165 170 175 Ser Ser Ala Leu AlaLeu Asp Asp His Ala Leu Leu His Asn Ser Ala 180 185 190 Asn Arg Trp GlnAsp Trp Asp Ile 195 200 <210> SEQ ID NO 21 <211> LENGTH: 704 <212> TYPE:DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:CDS <222> LOCATION: (157)...(597) <221> NAME/KEY: sig_peptide <222>LOCATION: (157)...(255) <221> NAME/KEY: mat_peptide <222> LOCATION:(256)...(597) <400> SEQUENCE: 21 aaaatcgctc ataaaatcgt gttttttgataccggtaaga tcgctgaaga aaacagcgct 60 aaagaatttt ttaaccaccc gaaatctcaaagagtgcaaa aatttttaga aactttccat 120 tttttaggga gctgttaaat aaagtttgctaaaaag atg gtt tta att tca aaa 174 Met Val Leu Ile Ser Lys -30 aga ggtgtt ttt atg aaa aca aac ggg ctt ttt aaa atg tgg ggg ctg 222 Arg Gly ValPhe Met Lys Thr Asn Gly Leu Phe Lys Met Trp Gly Leu -25 -20 -15 ttt ttagtt tta atc gct tta ctc ttt aac gca tgc tct gat agc cat 270 Phe Leu ValLeu Ile Ala Leu Leu Phe Asn Ala Cys Ser Asp Ser His -10 -5 1 5 aaa gaaaaa aag gac gct tta gaa gtc att aaa caa aga ggg gtt tta 318 Lys Glu LysLys Asp Ala Leu Glu Val Ile Lys Gln Arg Gly Val Leu 10 15 20 aaa gtg ggggtt ttt agc gat aag cct cct ttt gga tct gtg gat tct 366 Lys Val Gly ValPhe Ser Asp Lys Pro Pro Phe Gly Ser Val Asp Ser 25 30 35 aaa ggg aaa tatcaa ggc tat gat gtg atc atc gct aaa cgc atg gcc 414 Lys Gly Lys Tyr GlnGly Tyr Asp Val Ile Ile Ala Lys Arg Met Ala 40 45 50 ctt gat tta ttg ggcgat gaa aat aag att gag ttt ata ccc gta gaa 462 Leu Asp Leu Leu Gly AspGlu Asn Lys Ile Glu Phe Ile Pro Val Glu 55 60 65 gct tca gct agg gtg gaattt tta aaa gcc aat aaa gtg gat att atc 510 Ala Ser Ala Arg Val Glu PheLeu Lys Ala Asn Lys Val Asp Ile Ile 70 75 80 85 atg gct aat ttc acg cgcact aaa gaa aga gaa aaa gtc gtg gat ttc 558 Met Ala Asn Phe Thr Arg ThrLys Glu Arg Glu Lys Val Val Asp Phe 90 95 100 gct aat ccg tat atg aaagtc gct ttg ggg gtt gat ttc taaagatggg 607 Ala Asn Pro Tyr Met Lys ValAla Leu Gly Val Asp Phe 105 110 gtcattaaaa atatagaaga gttgaaggataaagagttga ttgtgaataa aggcacgaca 667 gcggattttt atttcactaa aaattaccccaatatca 704 <210> SEQ ID NO 22 <211> LENGTH: 147 <212> TYPE: PRT <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 22 Met Val Leu Ile Ser LysArg Gly Val Phe Met Lys Thr Asn Gly Leu 1 5 10 15 Phe Lys Met Trp GlyLeu Phe Leu Val Leu Ile Ala Leu Leu Phe Asn 20 25 30 Ala Cys Ser Asp SerHis Lys Glu Lys Lys Asp Ala Leu Glu Val Ile 35 40 45 Lys Gln Arg Gly ValLeu Lys Val Gly Val Phe Ser Asp Lys Pro Pro 50 55 60 Phe Gly Ser Val AspSer Lys Gly Lys Tyr Gln Gly Tyr Asp Val Ile 65 70 75 80 Ile Ala Lys ArgMet Ala Leu Asp Leu Leu Gly Asp Glu Asn Lys Ile 85 90 95 Glu Phe Ile ProVal Glu Ala Ser Ala Arg Val Glu Phe Leu Lys Ala 100 105 110 Asn Lys ValAsp Ile Ile Met Ala Asn Phe Thr Arg Thr Lys Glu Arg 115 120 125 Glu LysVal Val Asp Phe Ala Asn Pro Tyr Met Lys Val Ala Leu Gly 130 135 140 ValAsp Phe 145 <210> SEQ ID NO 23 <211> LENGTH: 1306 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY: CDS <222>LOCATION: (40)...(1266) <221> NAME/KEY: sig_peptide <222> LOCATION:(40)...(219) <221> NAME/KEY: mat_peptide <222> LOCATION: (220)...(1266)<400> SEQUENCE: 23 tttgacagct tatcatttgg caataaaaca ccaaaatga atg agttac aca aaa 54 Met Ser Tyr Thr Lys -60 aaa tac tca aca cca ccc aac cggcgt aaa atg caa aac att atc gct 102 Lys Tyr Ser Thr Pro Pro Asn Arg ArgLys Met Gln Asn Ile Ile Ala -55 -50 -45 -40 att aaa aga tcc tct aga gtcgac ctg cag gca tgc aag cta gct ttc 150 Ile Lys Arg Ser Ser Arg Val AspLeu Gln Ala Cys Lys Leu Ala Phe -35 -30 -25 gcg agc tcg aga tca ccc atgcaa ttt caa aaa acc tta ttt cct tta 198 Ala Ser Ser Arg Ser Pro Met GlnPhe Gln Lys Thr Leu Phe Pro Leu -20 -15 -10 ccc tta tta ttt tta tct tgttgt atc gct gaa gaa aat ggg gcg tat 246 Pro Leu Leu Phe Leu Ser Cys CysIle Ala Glu Glu Asn Gly Ala Tyr -5 1 5 gcg agc gtg ggg ttt gaa tat tccatt agt cat gcc gtt gag cat aat 294 Ala Ser Val Gly Phe Glu Tyr Ser IleSer His Ala Val Glu His Asn 10 15 20 25 aac cct ttt tta aat caa gaa cgcatc caa atc att tct aac gct caa 342 Asn Pro Phe Leu Asn Gln Glu Arg IleGln Ile Ile Ser Asn Ala Gln 30 35 40 aac aaa atc tat aaa ctc aat caa gtcaaa aat gaa atc aca agc atg 390 Asn Lys Ile Tyr Lys Leu Asn Gln Val LysAsn Glu Ile Thr Ser Met 45 50 55 caa aac acc ttt aat tac atc aac aac gcttta aaa aac aat gct aaa 438 Gln Asn Thr Phe Asn Tyr Ile Asn Asn Ala LeuLys Asn Asn Ala Lys 60 65 70 tta acc ccc act gaa atc caa gct gag aaa tactac ctc caa tcc acc 486 Leu Thr Pro Thr Glu Ile Gln Ala Glu Lys Tyr TyrLeu Gln Ser Thr 75 80 85 ctt caa aac att gaa aaa ata gtc aca ctt agc ggtggc gtt gca tct 534 Leu Gln Asn Ile Glu Lys Ile Val Thr Leu Ser Gly GlyVal Ala Ser 90 95 100 105 aac ccc aaa cta gtc caa gcg ttg gaa aaa atgcaa gaa ccc att act 582 Asn Pro Lys Leu Val Gln Ala Leu Glu Lys Met GlnGlu Pro Ile Thr 110 115 120 aac cct tta gaa tta gca gaa aac tta aga aattta gaa ttg caa ttt 630 Asn Pro Leu Glu Leu Ala Glu Asn Leu Arg Asn LeuGlu Leu Gln Phe 125 130 135 gct caa tct caa aac cgc atg ctt tct tct ttatct tct caa acc gct 678 Ala Gln Ser Gln Asn Arg Met Leu Ser Ser Leu SerSer Gln Thr Ala 140 145 150 caa att tca aat tct ttg aac gcg ctt gat cccagc tct tat tct aaa 726 Gln Ile Ser Asn Ser Leu Asn Ala Leu Asp Pro SerSer Tyr Ser Lys 155 160 165 aac att tca agc atg tct ggg gtg agt ttg agcgta ggg tat aag cat 774 Asn Ile Ser Ser Met Ser Gly Val Ser Leu Ser ValGly Tyr Lys His 170 175 180 185 ttc ttt act aag aaa aaa aat caa ggg tttcgc tat tac ttg ttt tat 822 Phe Phe Thr Lys Lys Lys Asn Gln Gly Phe ArgTyr Tyr Leu Phe Tyr 190 195 200 gac tat ggt tac act aac ttt ggt ttt gtgggt aat ggc ttt gat ggt 870 Asp Tyr Gly Tyr Thr Asn Phe Gly Phe Val GlyAsn Gly Phe Asp Gly 205 210 215 tta ggc aaa atg aat aac cac ctc tat gggctt gga ata aac tac ctt 918 Leu Gly Lys Met Asn Asn His Leu Tyr Gly LeuGly Ile Asn Tyr Leu 220 225 230 tat aat ttc att gat aat gca caa aaa cattcg agc gtg ggt ttt tat 966 Tyr Asn Phe Ile Asp Asn Ala Gln Lys His SerSer Val Gly Phe Tyr 235 240 245 gcg ggt ttt gct ttg gcg ggg aat tcg tgggta ggg aat ggt tta ggc 1014 Ala Gly Phe Ala Leu Ala Gly Asn Ser Trp ValGly Asn Gly Leu Gly 250 255 260 265 atg tgg gtg agc caa acg gat ttt atcaac aat tac ttg atg ggc tat 1062 Met Trp Val Ser Gln Thr Asp Phe Ile AsnAsn Tyr Leu Met Gly Tyr 270 275 280 caa gct aaa ata cac acg aac ttt ttccag atc cct ttg aat ttt ggg 1110 Gln Ala Lys Ile His Thr Asn Phe Phe GlnIle Pro Leu Asn Phe Gly 285 290 295 gtt cgt gtg aat gtc aat agg cat aacgga ttt gaa atg ggc cta aaa 1158 Val Arg Val Asn Val Asn Arg His Asn GlyPhe Glu Met Gly Leu Lys 300 305 310 atc cct tta gcg gtg aat tcc ttt tatgaa acg cat ggc aaa ggg tta 1206 Ile Pro Leu Ala Val Asn Ser Phe Tyr GluThr His Gly Lys Gly Leu 315 320 325 aac act tcc ctc ttt ttc aaa cgc cttgtg gtg ttt aat gtg agt tat 1254 Asn Thr Ser Leu Phe Phe Lys Arg Leu ValVal Phe Asn Val Ser Tyr 330 335 340 345 gtt tat agt ttt tagggggtaaatgccttcaa acgctctttt gattgaagaa 1306 Val Tyr Ser Phe <210> SEQ ID NO 24<211> LENGTH: 409 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 24 Met Ser Tyr Thr Lys Lys Tyr Ser Thr Pro Pro Asn ArgArg Lys Met 1 5 10 15 Gln Asn Ile Ile Ala Ile Lys Arg Ser Ser Arg ValAsp Leu Gln Ala 20 25 30 Cys Lys Leu Ala Phe Ala Ser Ser Arg Ser Pro MetGln Phe Gln Lys 35 40 45 Thr Leu Phe Pro Leu Pro Leu Leu Phe Leu Ser CysCys Ile Ala Glu 50 55 60 Glu Asn Gly Ala Tyr Ala Ser Val Gly Phe Glu TyrSer Ile Ser His 65 70 75 80 Ala Val Glu His Asn Asn Pro Phe Leu Asn GlnGlu Arg Ile Gln Ile 85 90 95 Ile Ser Asn Ala Gln Asn Lys Ile Tyr Lys LeuAsn Gln Val Lys Asn 100 105 110 Glu Ile Thr Ser Met Gln Asn Thr Phe AsnTyr Ile Asn Asn Ala Leu 115 120 125 Lys Asn Asn Ala Lys Leu Thr Pro ThrGlu Ile Gln Ala Glu Lys Tyr 130 135 140 Tyr Leu Gln Ser Thr Leu Gln AsnIle Glu Lys Ile Val Thr Leu Ser 145 150 155 160 Gly Gly Val Ala Ser AsnPro Lys Leu Val Gln Ala Leu Glu Lys Met 165 170 175 Gln Glu Pro Ile ThrAsn Pro Leu Glu Leu Ala Glu Asn Leu Arg Asn 180 185 190 Leu Glu Leu GlnPhe Ala Gln Ser Gln Asn Arg Met Leu Ser Ser Leu 195 200 205 Ser Ser GlnThr Ala Gln Ile Ser Asn Ser Leu Asn Ala Leu Asp Pro 210 215 220 Ser SerTyr Ser Lys Asn Ile Ser Ser Met Ser Gly Val Ser Leu Ser 225 230 235 240Val Gly Tyr Lys His Phe Phe Thr Lys Lys Lys Asn Gln Gly Phe Arg 245 250255 Tyr Tyr Leu Phe Tyr Asp Tyr Gly Tyr Thr Asn Phe Gly Phe Val Gly 260265 270 Asn Gly Phe Asp Gly Leu Gly Lys Met Asn Asn His Leu Tyr Gly Leu275 280 285 Gly Ile Asn Tyr Leu Tyr Asn Phe Ile Asp Asn Ala Gln Lys HisSer 290 295 300 Ser Val Gly Phe Tyr Ala Gly Phe Ala Leu Ala Gly Asn SerTrp Val 305 310 315 320 Gly Asn Gly Leu Gly Met Trp Val Ser Gln Thr AspPhe Ile Asn Asn 325 330 335 Tyr Leu Met Gly Tyr Gln Ala Lys Ile His ThrAsn Phe Phe Gln Ile 340 345 350 Pro Leu Asn Phe Gly Val Arg Val Asn ValAsn Arg His Asn Gly Phe 355 360 365 Glu Met Gly Leu Lys Ile Pro Leu AlaVal Asn Ser Phe Tyr Glu Thr 370 375 380 His Gly Lys Gly Leu Asn Thr SerLeu Phe Phe Lys Arg Leu Val Val 385 390 395 400 Phe Asn Val Ser Tyr ValTyr Ser Phe 405 <210> SEQ ID NO 25 <211> LENGTH: 999 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY: CDS<222> LOCATION: (25)...(942) <221> NAME/KEY: sig_peptide <222> LOCATION:(25)...(78) <221> NAME/KEY: mat_peptide <222> LOCATION: (79)...(942)<400> SEQUENCE: 25 tgacttagtt ttgtaaggaa tgag atg ata aag agt tgg actaaa aag tgg 51 Met Ile Lys Ser Trp Thr Lys Lys Trp -15 -10 ttt ttg atttta ttt tta atg gca agc tgt ttc agt tat ttg gtg gct 99 Phe Leu Ile LeuPhe Leu Met Ala Ser Cys Phe Ser Tyr Leu Val Ala -5 1 5 aca acc ggt gagaaa tat ttt aaa atg gct act caa gcc ttt aag aga 147 Thr Thr Gly Glu LysTyr Phe Lys Met Ala Thr Gln Ala Phe Lys Arg 10 15 20 ggg gat tac cat aaagcg gtg gct ttt tat aag agg agc tgt aat tta 195 Gly Asp Tyr His Lys AlaVal Ala Phe Tyr Lys Arg Ser Cys Asn Leu 25 30 35 agg gtg ggg gtt ggt tgcacg agt ttg ggc tct atg tat gaa gat ggc 243 Arg Val Gly Val Gly Cys ThrSer Leu Gly Ser Met Tyr Glu Asp Gly 40 45 50 55 gat ggc gtg gat cag aatatt cca aaa gcc gtt ttt tat tac aga aga 291 Asp Gly Val Asp Gln Asn IlePro Lys Ala Val Phe Tyr Tyr Arg Arg 60 65 70 ggg tgc aat ttg agg aat tatttg gct tgt gcg agt tta ggc tct atg 339 Gly Cys Asn Leu Arg Asn Tyr LeuAla Cys Ala Ser Leu Gly Ser Met 75 80 85 tat gaa gat ggc gat ggc gtt caaaaa aac ctt cca aag gct atc tat 387 Tyr Glu Asp Gly Asp Gly Val Gln LysAsn Leu Pro Lys Ala Ile Tyr 90 95 100 tat tat agg aga ggg tgc cac ttaaag ggt ggg gtg agc tgc ggg agt 435 Tyr Tyr Arg Arg Gly Cys His Leu LysGly Gly Val Ser Cys Gly Ser 105 110 115 tta ggt ttt atg tat ttt aat ggcacg ggc gtt aag caa aat tat gcc 483 Leu Gly Phe Met Tyr Phe Asn Gly ThrGly Val Lys Gln Asn Tyr Ala 120 125 130 135 aaa gcc ctt tct ctt tct caatac gct tgc agt ttg aat tat ggc att 531 Lys Ala Leu Ser Leu Ser Gln TyrAla Cys Ser Leu Asn Tyr Gly Ile 140 145 150 agt tgt aac ttt gta ggg tatatg tat agg agc gcc aaa ggc gta cag 579 Ser Cys Asn Phe Val Gly Tyr MetTyr Arg Ser Ala Lys Gly Val Gln 155 160 165 aag gat ttg aaa aaa gcc cttacg aat ttt aaa aga ggg tgc cat tta 627 Lys Asp Leu Lys Lys Ala Leu ThrAsn Phe Lys Arg Gly Cys His Leu 170 175 180 aaa gac gga gcg agc tgt gtgagc ttg gga tac atg tat gaa gtc ggc 675 Lys Asp Gly Ala Ser Cys Val SerLeu Gly Tyr Met Tyr Glu Val Gly 185 190 195 atg tat gtc aga caa aat gaagag caa gcc ttg aat ctt tat aaa aag 723 Met Tyr Val Arg Gln Asn Glu GluGln Ala Leu Asn Leu Tyr Lys Lys 200 205 210 215 ggt tgt cat tta aaa gaaggg agc ggt tgc cat aat gtg gcg gtg atg 771 Gly Cys His Leu Lys Glu GlySer Gly Cys His Asn Val Ala Val Met 220 225 230 tat tac acg ggt aag ggtgct tca aag gat tta gat aaa gcc att tcg 819 Tyr Tyr Thr Gly Lys Gly AlaSer Lys Asp Leu Asp Lys Ala Ile Ser 235 240 245 tat tat aag aaa ggt tgcact cta ggc ttt agt ggt agc tgt aag gtg 867 Tyr Tyr Lys Lys Gly Cys ThrLeu Gly Phe Ser Gly Ser Cys Lys Val 250 255 260 tta gaa gaa gtg att ggcaag aag tct gat gat ttg caa gat gat gcg 915 Leu Glu Glu Val Ile Gly LysLys Ser Asp Asp Leu Gln Asp Asp Ala 265 270 275 caa aac gac acg caa gatgat acg caa taagccagag tttaggggct 962 Gln Asn Asp Thr Gln Asp Asp ThrGln 280 285 aatgattaaa actcatctta tagaaatctt tctattc 999 <210> SEQ ID NO26 <211> LENGTH: 306 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 26 Met Ile Lys Ser Trp Thr Lys Lys Trp Phe Leu Ile LeuPhe Leu Met 1 5 10 15 Ala Ser Cys Phe Ser Tyr Leu Val Ala Thr Thr GlyGlu Lys Tyr Phe 20 25 30 Lys Met Ala Thr Gln Ala Phe Lys Arg Gly Asp TyrHis Lys Ala Val 35 40 45 Ala Phe Tyr Lys Arg Ser Cys Asn Leu Arg Val GlyVal Gly Cys Thr 50 55 60 Ser Leu Gly Ser Met Tyr Glu Asp Gly Asp Gly ValAsp Gln Asn Ile 65 70 75 80 Pro Lys Ala Val Phe Tyr Tyr Arg Arg Gly CysAsn Leu Arg Asn Tyr 85 90 95 Leu Ala Cys Ala Ser Leu Gly Ser Met Tyr GluAsp Gly Asp Gly Val 100 105 110 Gln Lys Asn Leu Pro Lys Ala Ile Tyr TyrTyr Arg Arg Gly Cys His 115 120 125 Leu Lys Gly Gly Val Ser Cys Gly SerLeu Gly Phe Met Tyr Phe Asn 130 135 140 Gly Thr Gly Val Lys Gln Asn TyrAla Lys Ala Leu Ser Leu Ser Gln 145 150 155 160 Tyr Ala Cys Ser Leu AsnTyr Gly Ile Ser Cys Asn Phe Val Gly Tyr 165 170 175 Met Tyr Arg Ser AlaLys Gly Val Gln Lys Asp Leu Lys Lys Ala Leu 180 185 190 Thr Asn Phe LysArg Gly Cys His Leu Lys Asp Gly Ala Ser Cys Val 195 200 205 Ser Leu GlyTyr Met Tyr Glu Val Gly Met Tyr Val Arg Gln Asn Glu 210 215 220 Glu GlnAla Leu Asn Leu Tyr Lys Lys Gly Cys His Leu Lys Glu Gly 225 230 235 240Ser Gly Cys His Asn Val Ala Val Met Tyr Tyr Thr Gly Lys Gly Ala 245 250255 Ser Lys Asp Leu Asp Lys Ala Ile Ser Tyr Tyr Lys Lys Gly Cys Thr 260265 270 Leu Gly Phe Ser Gly Ser Cys Lys Val Leu Glu Glu Val Ile Gly Lys275 280 285 Lys Ser Asp Asp Leu Gln Asp Asp Ala Gln Asn Asp Thr Gln AspAsp 290 295 300 Thr Gln 305 <210> SEQ ID NO 27 <211> LENGTH: 805 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: CDS <222> LOCATION: (130)...(642) <221> NAME/KEY: sig_peptide<222> LOCATION: (130)...(192) <221> NAME/KEY: mat_peptide <222>LOCATION: (193)...(642) <400> SEQUENCE: 27 tagtgagagt cgcccaattccaatgatgga aagcatgcaa gtctttcatg atatatcgcc 60 atttttaaaa agttttgagactataaccct taaaaaacta tcgcaacaaa tcaattcata 120 ggaacaagc atg ttt tttaaa act tat caa aaa tta ctg ggt gca agc tgt 171 Met Phe Phe Lys Thr TyrGln Lys Leu Leu Gly Ala Ser Cys -20 -15 -10 ttg gcg tta tat tta gca ggctgt ggg ggc gac agt ggc gaa cca cta 219 Leu Ala Leu Tyr Leu Ala Gly CysGly Gly Asp Ser Gly Glu Pro Leu -5 1 5 gtt ggg att gaa aaa aat agc ttcaat tct acc ttg aaa atc att tct 267 Val Gly Ile Glu Lys Asn Ser Phe AsnSer Thr Leu Lys Ile Ile Ser 10 15 20 25 aaa acc gac aac ata gaa atc caagac ttg aag ctc aat cgt gag aat 315 Lys Thr Asp Asn Ile Glu Ile Gln AspLeu Lys Leu Asn Arg Glu Asn 30 35 40 tgc gag cat gat gaa aat ttc ttg gtaaag tta ata caa gaa aca gcc 363 Cys Glu His Asp Glu Asn Phe Leu Val LysLeu Ile Gln Glu Thr Ala 45 50 55 aat aca tac ctg ttt gca tca gaa aaa gaaaaa gcg att aaa aac cac 411 Asn Thr Tyr Leu Phe Ala Ser Glu Lys Glu LysAla Ile Lys Asn His 60 65 70 caa gca aaa atc gca aga ctt caa aaa aga aggagc ggt tgc cat aat 459 Gln Ala Lys Ile Ala Arg Leu Gln Lys Arg Arg SerGly Cys His Asn 75 80 85 gtg gcg gtg atg tat tac acg ggt aag ggt gct tcaaag gat tta gat 507 Val Ala Val Met Tyr Tyr Thr Gly Lys Gly Ala Ser LysAsp Leu Asp 90 95 100 105 aaa gcc att tcg tat tat aag aaa ggt tgc actcta ggc ttt agt ggt 555 Lys Ala Ile Ser Tyr Tyr Lys Lys Gly Cys Thr LeuGly Phe Ser Gly 110 115 120 agc tgt aag gtg tta gaa gaa gtg att ggc aagaag tct gat gat ttg 603 Ser Cys Lys Val Leu Glu Glu Val Ile Gly Lys LysSer Asp Asp Leu 125 130 135 caa gat gat gcg caa aac gac acg caa gat gatacg caa taagccagag 652 Gln Asp Asp Ala Gln Asn Asp Thr Gln Asp Asp ThrGln 140 145 150 tttaggggct aatgattaaa actcatctta tagaaatctt tctattctcttgctatcaaa 712 tagggattga gcgtctctat tgatgggtat tgagattaaa aacctgcaaatctagggttt 772 ggatttattc tcatatggat aataaaaata ctc 805 <210> SEQ ID NO28 <211> LENGTH: 171 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 28 Met Phe Phe Lys Thr Tyr Gln Lys Leu Leu Gly Ala SerCys Leu Ala 1 5 10 15 Leu Tyr Leu Ala Gly Cys Gly Gly Asp Ser Gly GluPro Leu Val Gly 20 25 30 Ile Glu Lys Asn Ser Phe Asn Ser Thr Leu Lys IleIle Ser Lys Thr 35 40 45 Asp Asn Ile Glu Ile Gln Asp Leu Lys Leu Asn ArgGlu Asn Cys Glu 50 55 60 His Asp Glu Asn Phe Leu Val Lys Leu Ile Gln GluThr Ala Asn Thr 65 70 75 80 Tyr Leu Phe Ala Ser Glu Lys Glu Lys Ala IleLys Asn His Gln Ala 85 90 95 Lys Ile Ala Arg Leu Gln Lys Arg Arg Ser GlyCys His Asn Val Ala 100 105 110 Val Met Tyr Tyr Thr Gly Lys Gly Ala SerLys Asp Leu Asp Lys Ala 115 120 125 Ile Ser Tyr Tyr Lys Lys Gly Cys ThrLeu Gly Phe Ser Gly Ser Cys 130 135 140 Lys Val Leu Glu Glu Val Ile GlyLys Lys Ser Asp Asp Leu Gln Asp 145 150 155 160 Asp Ala Gln Asn Asp ThrGln Asp Asp Thr Gln 165 170 <210> SEQ ID NO 29 <211> LENGTH: 1101 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: CDS <222> LOCATION: (40)...(1026) <221> NAME/KEY: sig_peptide<222> LOCATION: (40)...(99) <221> NAME/KEY: mat_peptide <222> LOCATION:(100)...(1026) <400> SEQUENCE: 29 ggttataccg aaaaaacaat atgaaatcaaggagtttgt atg caa cag cgt cat 54 Met Gln Gln Arg His -20 tta ggc cct ttaaaa gtg ggt gca tta gct cta ggg tgc atg ggc atg 102 Leu Gly Pro Leu LysVal Gly Ala Leu Ala Leu Gly Cys Met Gly Met -15 -10 -5 1 act tat ggg tatggg gaa gtc cat gat aaa aag cag atg gtt aaa ctt 150 Thr Tyr Gly Tyr GlyGlu Val His Asp Lys Lys Gln Met Val Lys Leu 5 10 15 atc cat aag gct ttggaa ttg ggt att aac ttt ttt gac act gca gag 198 Ile His Lys Ala Leu GluLeu Gly Ile Asn Phe Phe Asp Thr Ala Glu 20 25 30 gct tat ggg gaa gat aatgaa aag ctt tta gcg aag cga tca agc ctt 246 Ala Tyr Gly Glu Asp Asn GluLys Leu Leu Ala Lys Arg Ser Ser Leu 35 40 45 att aaa gac aag gtt gtg gtagcg agc aag ttt ggg att tac tac gca 294 Ile Lys Asp Lys Val Val Val AlaSer Lys Phe Gly Ile Tyr Tyr Ala 50 55 60 65 gat cct aat gac aaa tac gcaacc atg ttt tta gac tcc agt tct aac 342 Asp Pro Asn Asp Lys Tyr Ala ThrMet Phe Leu Asp Ser Ser Ser Asn 70 75 80 cgc att aag agt gcc att gaa gggagt ttg aaa cgc tta aaa gta gaa 390 Arg Ile Lys Ser Ala Ile Glu Gly SerLeu Lys Arg Leu Lys Val Glu 85 90 95 tgc att gat tta tac tac caa cac cgcatg gat act aac acg ccc ata 438 Cys Ile Asp Leu Tyr Tyr Gln His Arg MetAsp Thr Asn Thr Pro Ile 100 105 110 gaa gaa gtg gca gaa gtt atg caa gctctt att aaa gaa gga aaa att 486 Glu Glu Val Ala Glu Val Met Gln Ala LeuIle Lys Glu Gly Lys Ile 115 120 125 aaa gct tgg ggg atg agt gag gca gggtta tct agc atc caa aaa gcc 534 Lys Ala Trp Gly Met Ser Glu Ala Gly LeuSer Ser Ile Gln Lys Ala 130 135 140 145 cat caa att tgc cct tta agc gcgttg cag agc gaa tat tcc ttg tgg 582 His Gln Ile Cys Pro Leu Ser Ala LeuGln Ser Glu Tyr Ser Leu Trp 150 155 160 tgg cgc gaa cct gaa aaa gag atttta ggt ttt tta gaa aaa gaa aaa 630 Trp Arg Glu Pro Glu Lys Glu Ile LeuGly Phe Leu Glu Lys Glu Lys 165 170 175 att ggc ttt gtc gct ttt tcg cctttg ggt aag ggg ttt tta ggc gcg 678 Ile Gly Phe Val Ala Phe Ser Pro LeuGly Lys Gly Phe Leu Gly Ala 180 185 190 aaa ttt gaa aaa aat gct acc ttcgct agt gaa gat ttt aga agc gtt 726 Lys Phe Glu Lys Asn Ala Thr Phe AlaSer Glu Asp Phe Arg Ser Val 195 200 205 tct cct agg ttt aat caa gaa aatcta gcc aaa aat tac gtc ttg gtg 774 Ser Pro Arg Phe Asn Gln Glu Asn LeuAla Lys Asn Tyr Val Leu Val 210 215 220 225 gaa tta atc caa gat cat gcacac gct aaa ggc gtt aca cca gcc caa 822 Glu Leu Ile Gln Asp His Ala HisAla Lys Gly Val Thr Pro Ala Gln 230 235 240 ctg gct ctc tcg tgg att ttgcac acg caa aaa atc att gtc cct ctc 870 Leu Ala Leu Ser Trp Ile Leu HisThr Gln Lys Ile Ile Val Pro Leu 245 250 255 ttt ggc acc acc aaa gaa tccagg ctc ata gaa aat ata ggg gct ttg 918 Phe Gly Thr Thr Lys Glu Ser ArgLeu Ile Glu Asn Ile Gly Ala Leu 260 265 270 cag gtt tct tgg agt caa aaagaa ttg gag att ttt caa aaa gaa ttg 966 Gln Val Ser Trp Ser Gln Lys GluLeu Glu Ile Phe Gln Lys Glu Leu 275 280 285 act gca atc aaa ata gaa ggggcc cgc tac cct gaa aga atc aat gaa 1014 Thr Ala Ile Lys Ile Glu Gly AlaArg Tyr Pro Glu Arg Ile Asn Glu 290 295 300 305 atg gtg aat caataaaagtatt gggtatttat aattgcattg gctcttttaa 1066 Met Val Asn Glnaagagattga gcgttatttc ctgtttgtca gtgtg 1101 <210> SEQ ID NO 30 <211>LENGTH: 329 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 30 Met Gln Gln Arg His Leu Gly Pro Leu Lys Val Gly Ala Leu AlaLeu 1 5 10 15 Gly Cys Met Gly Met Thr Tyr Gly Tyr Gly Glu Val His AspLys Lys 20 25 30 Gln Met Val Lys Leu Ile His Lys Ala Leu Glu Leu Gly IleAsn Phe 35 40 45 Phe Asp Thr Ala Glu Ala Tyr Gly Glu Asp Asn Glu Lys LeuLeu Ala 50 55 60 Lys Arg Ser Ser Leu Ile Lys Asp Lys Val Val Val Ala SerLys Phe 65 70 75 80 Gly Ile Tyr Tyr Ala Asp Pro Asn Asp Lys Tyr Ala ThrMet Phe Leu 85 90 95 Asp Ser Ser Ser Asn Arg Ile Lys Ser Ala Ile Glu GlySer Leu Lys 100 105 110 Arg Leu Lys Val Glu Cys Ile Asp Leu Tyr Tyr GlnHis Arg Met Asp 115 120 125 Thr Asn Thr Pro Ile Glu Glu Val Ala Glu ValMet Gln Ala Leu Ile 130 135 140 Lys Glu Gly Lys Ile Lys Ala Trp Gly MetSer Glu Ala Gly Leu Ser 145 150 155 160 Ser Ile Gln Lys Ala His Gln IleCys Pro Leu Ser Ala Leu Gln Ser 165 170 175 Glu Tyr Ser Leu Trp Trp ArgGlu Pro Glu Lys Glu Ile Leu Gly Phe 180 185 190 Leu Glu Lys Glu Lys IleGly Phe Val Ala Phe Ser Pro Leu Gly Lys 195 200 205 Gly Phe Leu Gly AlaLys Phe Glu Lys Asn Ala Thr Phe Ala Ser Glu 210 215 220 Asp Phe Arg SerVal Ser Pro Arg Phe Asn Gln Glu Asn Leu Ala Lys 225 230 235 240 Asn TyrVal Leu Val Glu Leu Ile Gln Asp His Ala His Ala Lys Gly 245 250 255 ValThr Pro Ala Gln Leu Ala Leu Ser Trp Ile Leu His Thr Gln Lys 260 265 270Ile Ile Val Pro Leu Phe Gly Thr Thr Lys Glu Ser Arg Leu Ile Glu 275 280285 Asn Ile Gly Ala Leu Gln Val Ser Trp Ser Gln Lys Glu Leu Glu Ile 290295 300 Phe Gln Lys Glu Leu Thr Ala Ile Lys Ile Glu Gly Ala Arg Tyr Pro305 310 315 320 Glu Arg Ile Asn Glu Met Val Asn Gln 325 <210> SEQ ID NO31 <211> LENGTH: 495 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (1)...(492) <221>NAME/KEY: sig_peptide <222> LOCATION: (1)...(105) <221> NAME/KEY:mat_peptide <222> LOCATION: (106)...(492) <400> SEQUENCE: 31 atg gta tttgac aga aca atc agc gta aga gaa aaa aaa gcg gct aaa 48 Met Val Phe AspArg Thr Ile Ser Val Arg Glu Lys Lys Ala Ala Lys -35 -30 -25 -20 acg cttggg att gtg ggg atc gtc ttt ttt att ttg ttt ggc atc gta 96 Thr Leu GlyIle Val Gly Ile Val Phe Phe Ile Leu Phe Gly Ile Val -15 -10 -5 ata agcggg gtg gct ttt caa aaa gag tgg gtg caa caa ttg gat tta 144 Ile Ser GlyVal Ala Phe Gln Lys Glu Trp Val Gln Gln Leu Asp Leu 1 5 10 ttt ttt atagac ttg atc cac aac cct gcc ccc att caa ggg agc gcg 192 Phe Phe Ile AspLeu Ile His Asn Pro Ala Pro Ile Gln Gly Ser Ala 15 20 25 tgg ctt tct ttcgtg ttt ttt agc aca tgg ttt gcg caa agc aag ctc 240 Trp Leu Ser Phe ValPhe Phe Ser Thr Trp Phe Ala Gln Ser Lys Leu 30 35 40 45 acc act cct atagcc tta ctc att ggc ttg tgg ttt ggg ttt caa aaa 288 Thr Thr Pro Ile AlaLeu Leu Ile Gly Leu Trp Phe Gly Phe Gln Lys 50 55 60 cgc atc gct tta ggggtg tgg ttt ttc ttt agc atc tta tta ggt gaa 336 Arg Ile Ala Leu Gly ValTrp Phe Phe Phe Ser Ile Leu Leu Gly Glu 65 70 75 ttc acc tta aaa tcc cttaag ctt tta gtg gcg cgc cca cgg cct gta 384 Phe Thr Leu Lys Ser Leu LysLeu Leu Val Ala Arg Pro Arg Pro Val 80 85 90 acc aat ggc gaa ttg gtt tttgca cat ggc ttt agt ttc ccc agc ggg 432 Thr Asn Gly Glu Leu Val Phe AlaHis Gly Phe Ser Phe Pro Ser Gly 95 100 105 cat gct tta gct tcc agc gctttt tta cgg ctc ttt ggc gtt tgt ttg 480 His Ala Leu Ala Ser Ser Ala PheLeu Arg Leu Phe Gly Val Cys Leu 110 115 120 125 tta tgc tat tcc taa 495Leu Cys Tyr Ser <210> SEQ ID NO 32 <211> LENGTH: 164 <212> TYPE: PRT<213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 32 Met Val Phe AspArg Thr Ile Ser Val Arg Glu Lys Lys Ala Ala Lys 1 5 10 15 Thr Leu GlyIle Val Gly Ile Val Phe Phe Ile Leu Phe Gly Ile Val 20 25 30 Ile Ser GlyVal Ala Phe Gln Lys Glu Trp Val Gln Gln Leu Asp Leu 35 40 45 Phe Phe IleAsp Leu Ile His Asn Pro Ala Pro Ile Gln Gly Ser Ala 50 55 60 Trp Leu SerPhe Val Phe Phe Ser Thr Trp Phe Ala Gln Ser Lys Leu 65 70 75 80 Thr ThrPro Ile Ala Leu Leu Ile Gly Leu Trp Phe Gly Phe Gln Lys 85 90 95 Arg IleAla Leu Gly Val Trp Phe Phe Phe Ser Ile Leu Leu Gly Glu 100 105 110 PheThr Leu Lys Ser Leu Lys Leu Leu Val Ala Arg Pro Arg Pro Val 115 120 125Thr Asn Gly Glu Leu Val Phe Ala His Gly Phe Ser Phe Pro Ser Gly 130 135140 His Ala Leu Ala Ser Ser Ala Phe Leu Arg Leu Phe Gly Val Cys Leu 145150 155 160 Leu Cys Tyr Ser <210> SEQ ID NO 33 <211> LENGTH: 569 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: CDS <222> LOCATION: (1)...(516) <221> NAME/KEY: sig_peptide<222> LOCATION: (1)...(57) <221> NAME/KEY: mat_peptide <222> LOCATION:(58)...(516) <400> SEQUENCE: 33 atg ttg aaa ttt aaa tat ggt ttg att tatatc gcg ctc att ata gga 48 Met Leu Lys Phe Lys Tyr Gly Leu Ile Tyr IleAla Leu Ile Ile Gly -15 -10 -5 ctt caa gcg aca gat tat gac aat tta gaagaa gaa aac caa caa tta 96 Leu Gln Ala Thr Asp Tyr Asp Asn Leu Glu GluGlu Asn Gln Gln Leu 1 5 10 gac gaa aaa ata aac cat tta aag caa cag cttacc gaa aaa ggg gtt 144 Asp Glu Lys Ile Asn His Leu Lys Gln Gln Leu ThrGlu Lys Gly Val 15 20 25 tcg ccc aaa gag atg gat aag gat aag ttt gaa gaagaa tat tta gag 192 Ser Pro Lys Glu Met Asp Lys Asp Lys Phe Glu Glu GluTyr Leu Glu 30 35 40 45 cga act tac cca aag att tct tca aag aaa aga aaaaaa tta ctc aaa 240 Arg Thr Tyr Pro Lys Ile Ser Ser Lys Lys Arg Lys LysLeu Leu Lys 50 55 60 tct ttc tcc ata gcc gat gat aag agt ggg gtt ttt ttaggg ggt ggg 288 Ser Phe Ser Ile Ala Asp Asp Lys Ser Gly Val Phe Leu GlyGly Gly 65 70 75 tat gct tat ggg gga ttt aat ctt tct tat caa ggg gag atgtta gac 336 Tyr Ala Tyr Gly Gly Phe Asn Leu Ser Tyr Gln Gly Glu Met LeuAsp 80 85 90 aaa tat ggt gcg aat gcc cct agt gtg ttt aaa aac aat att aagatt 384 Lys Tyr Gly Ala Asn Ala Pro Ser Val Phe Lys Asn Asn Ile Lys Ile95 100 105 aac gct cct gtt tct atg att agc gtt aaa ttc ggg tat caa aaatac 432 Asn Ala Pro Val Ser Met Ile Ser Val Lys Phe Gly Tyr Gln Lys Tyr110 115 120 125 ttt gtg cct tat ttt ggg aca cga ttt tat ggg gat tta ttgctt ggg 480 Phe Val Pro Tyr Phe Gly Thr Arg Phe Tyr Gly Asp Leu Leu LeuGly 130 135 140 ggt gga gcg tta aaa agg atg caa gca agc aac ctgtaggctcgtt 526 Gly Gly Ala Leu Lys Arg Met Gln Ala Ser Asn Leu 145 150tatttatgtt tttaggggct atgaatacgg atttattgtt tga 569 <210> SEQ ID NO 34<211> LENGTH: 172 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 34 Met Leu Lys Phe Lys Tyr Gly Leu Ile Tyr Ile Ala LeuIle Ile Gly 1 5 10 15 Leu Gln Ala Thr Asp Tyr Asp Asn Leu Glu Glu GluAsn Gln Gln Leu 20 25 30 Asp Glu Lys Ile Asn His Leu Lys Gln Gln Leu ThrGlu Lys Gly Val 35 40 45 Ser Pro Lys Glu Met Asp Lys Asp Lys Phe Glu GluGlu Tyr Leu Glu 50 55 60 Arg Thr Tyr Pro Lys Ile Ser Ser Lys Lys Arg LysLys Leu Leu Lys 65 70 75 80 Ser Phe Ser Ile Ala Asp Asp Lys Ser Gly ValPhe Leu Gly Gly Gly 85 90 95 Tyr Ala Tyr Gly Gly Phe Asn Leu Ser Tyr GlnGly Glu Met Leu Asp 100 105 110 Lys Tyr Gly Ala Asn Ala Pro Ser Val PheLys Asn Asn Ile Lys Ile 115 120 125 Asn Ala Pro Val Ser Met Ile Ser ValLys Phe Gly Tyr Gln Lys Tyr 130 135 140 Phe Val Pro Tyr Phe Gly Thr ArgPhe Tyr Gly Asp Leu Leu Leu Gly 145 150 155 160 Gly Gly Ala Leu Lys ArgMet Gln Ala Ser Asn Leu 165 170 <210> SEQ ID NO 35 <211> LENGTH: 1416<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: CDS <222> LOCATION: (315)...(917) <221> NAME/KEY: sig_peptide<222> LOCATION: (315)...(389) <221> NAME/KEY: mat_peptide <222>LOCATION: (390)...(917) <400> SEQUENCE: 35 gtattaccgc ctttgagtgagctgatacga attagatcat taaaggttcc ttttggagcc 60 tttttttttg aattctcatgtttgacagct tatcatttgg caataaaaca ccaaaatgaa 120 tgagttacac aaaaaaatactcaaacacca cccaaccggc gtaaaatgca aaacattatc 180 gctattaaaa gatcctctagagtcgacctg caggcatgca agctagcttt cgcgagctcg 240 agatcatttt agccataaaaagcttatgtt ttcattaaaa atgttatgat acgctcaaat 300 agtcaagcaa aaaa atg tcaatt aaa agg gtt aga ttg aaa ata ttc gtt 350 Met Ser Ile Lys Arg Val ArgLeu Lys Ile Phe Val -25 -20 -15 ctg ttg atg tcg gta att tta gga ata tcatta aca ggt tgc ata ggc 398 Leu Leu Met Ser Val Ile Leu Gly Ile Ser LeuThr Gly Cys Ile Gly -10 -5 1 tat cgt atg gac tta gaa cat ttt aac acg ctctat tat gaa gaa agc 446 Tyr Arg Met Asp Leu Glu His Phe Asn Thr Leu TyrTyr Glu Glu Ser 5 10 15 cct aaa caa gct tat gaa tat tct aaa caa ttc actaag aaa aaa aag 494 Pro Lys Gln Ala Tyr Glu Tyr Ser Lys Gln Phe Thr LysLys Lys Lys 20 25 30 35 aac gct ctt tta tgg gac ttg caa aac ggc ttg agcgct tta tac gcc 542 Asn Ala Leu Leu Trp Asp Leu Gln Asn Gly Leu Ser AlaLeu Tyr Ala 40 45 50 aga gat tac cag act tct tta ggg gtg tta gat caa gccgag caa cgc 590 Arg Asp Tyr Gln Thr Ser Leu Gly Val Leu Asp Gln Ala GluGln Arg 55 60 65 ttt gat aaa acc caa agc gct ttc aca aga ggg gct ggt tatgtg ggc 638 Phe Asp Lys Thr Gln Ser Ala Phe Thr Arg Gly Ala Gly Tyr ValGly 70 75 80 gct acc atg att aat gat aac gtg cgc gct tat ggg ggg aat atttat 686 Ala Thr Met Ile Asn Asp Asn Val Arg Ala Tyr Gly Gly Asn Ile Tyr85 90 95 gag ggc gtt tta atc aat tat tac aaa gcg ata gac tac atg ctt tta734 Glu Gly Val Leu Ile Asn Tyr Tyr Lys Ala Ile Asp Tyr Met Leu Leu 100105 110 115 aac gat agc gcg aaa gct agg gtg caa ttc aac cgc gcg aac gaacgc 782 Asn Asp Ser Ala Lys Ala Arg Val Gln Phe Asn Arg Ala Asn Glu Arg120 125 130 cag cgc agg gct aaa gaa ttt tat tat gag gaa gtg caa aaa gccatt 830 Gln Arg Arg Ala Lys Glu Phe Tyr Tyr Glu Glu Val Gln Lys Ala Ile135 140 145 aaa gag atc gat tct agc aaa aag cac aat att aat atg gaa cgctct 878 Lys Glu Ile Asp Ser Ser Lys Lys His Asn Ile Asn Met Glu Arg Ser150 155 160 agg gct aga agt gag cga gat ttt aaa caa cac tta ttctaatttagac 927 Arg Ala Arg Ser Glu Arg Asp Phe Lys Gln His Leu Phe 165170 175 aaatacgaag cttatcaagg cttgcttaac ccagcggttt cgtatctttcagggttgttt 987 tacgctttaa atggggataa gaataagggg ttaggctatc ttaatgaagcctacgggatc 1047 agtcaaagcc cttttgtagc ccaagacttg gtttttttta aaaaccctaataggagtcat 1107 ttcacttgga tcatcattga agatggtaaa gagccgcaaa aaagccaatttaaaattgat 1167 gtgcctattt ttatgattga ttcggtttat aacgtgagta tagccttgcccaagctagaa 1227 aaaggggaag cgttttatca aaatttcact cttaaagatg gagaaaaagtaacgcccttt 1287 gacactttag cctcaataga tgcggtggtc gctagcgaat ttaggaagcagttaccctat 1347 attatcacta gagccattct atcggctact tttagaggtg ggcatgcaagcggtagcgaa 1407 ttattattt 1416 <210> SEQ ID NO 36 <211> LENGTH: 201<212> TYPE: PRT <213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 36Met Ser Ile Lys Arg Val Arg Leu Lys Ile Phe Val Leu Leu Met Ser 1 5 1015 Val Ile Leu Gly Ile Ser Leu Thr Gly Cys Ile Gly Tyr Arg Met Asp 20 2530 Leu Glu His Phe Asn Thr Leu Tyr Tyr Glu Glu Ser Pro Lys Gln Ala 35 4045 Tyr Glu Tyr Ser Lys Gln Phe Thr Lys Lys Lys Lys Asn Ala Leu Leu 50 5560 Trp Asp Leu Gln Asn Gly Leu Ser Ala Leu Tyr Ala Arg Asp Tyr Gln 65 7075 80 Thr Ser Leu Gly Val Leu Asp Gln Ala Glu Gln Arg Phe Asp Lys Thr 8590 95 Gln Ser Ala Phe Thr Arg Gly Ala Gly Tyr Val Gly Ala Thr Met Ile100 105 110 Asn Asp Asn Val Arg Ala Tyr Gly Gly Asn Ile Tyr Glu Gly ValLeu 115 120 125 Ile Asn Tyr Tyr Lys Ala Ile Asp Tyr Met Leu Leu Asn AspSer Ala 130 135 140 Lys Ala Arg Val Gln Phe Asn Arg Ala Asn Glu Arg GlnArg Arg Ala 145 150 155 160 Lys Glu Phe Tyr Tyr Glu Glu Val Gln Lys AlaIle Lys Glu Ile Asp 165 170 175 Ser Ser Lys Lys His Asn Ile Asn Met GluArg Ser Arg Ala Arg Ser 180 185 190 Glu Arg Asp Phe Lys Gln His Leu Phe195 200 <210> SEQ ID NO 37 <211> LENGTH: 738 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY: CDS <222>LOCATION: (201)...(731) <221> NAME/KEY: sig_peptide <222> LOCATION:(201)...(263) <221> NAME/KEY: mat_peptide <222> LOCATION: (264)...(731)<400> SEQUENCE: 37 cgcctgattg ttcctttccc tgctatggaa aaagcgaaataaaaacaaga aagtaaccta 60 tgatttcgct cgtttgatgg atggggctaa agaagtcaaatgctctgaat tcgctagcgt 120 gatgattgaa aacatgtgaa agagcgtttt ttaggctctggtatttgaat gcgattatta 180 ggctaatact atcataagga atg aag ttg ata aaa tttgtg cgt aat gtg gtt 233 Met Lys Leu Ile Lys Phe Val Arg Asn Val Val -20-15 tta ttc att tta aca gcg atc ttt tta gca ctc atg ctt tta gtg agc 281Leu Phe Ile Leu Thr Ala Ile Phe Leu Ala Leu Met Leu Leu Val Ser -10 -5 15 tat tgc atg ccc cat tat agc gtg gct gtc att agc ggg gtg gaa gtc 329Tyr Cys Met Pro His Tyr Ser Val Ala Val Ile Ser Gly Val Glu Val 10 15 20caa aga atg aat gaa aat gca cgc cca aat aat aag gaa gta aaa acc 377 GlnArg Met Asn Glu Asn Ala Arg Pro Asn Asn Lys Glu Val Lys Thr 25 30 35 ctagct aga gat gtc tat ttt gtg caa act tac gac cct aag gat caa 425 Leu AlaArg Asp Val Tyr Phe Val Gln Thr Tyr Asp Pro Lys Asp Gln 40 45 50 aaa agcgta acc gtc tat cgt aac gaa gac acg cgc ttt ggc ttc cct 473 Lys Ser ValThr Val Tyr Arg Asn Glu Asp Thr Arg Phe Gly Phe Pro 55 60 65 70 ttt tatttt aag ttt aat tcg gct gat att tca gcc ctc gct caa agt 521 Phe Tyr PheLys Phe Asn Ser Ala Asp Ile Ser Ala Leu Ala Gln Ser 75 80 85 tta gtc aaccag caa gtg gaa gtg caa tac tat ggc tgg cgg atc aat 569 Leu Val Asn GlnGln Val Glu Val Gln Tyr Tyr Gly Trp Arg Ile Asn 90 95 100 ttg ttt aacatg ttc cct aat gtg att ttt tta aag ccc tta aaa gag 617 Leu Phe Asn MetPhe Pro Asn Val Ile Phe Leu Lys Pro Leu Lys Glu 105 110 115 agt gct gagatg tca aaa ccc att ttt agc tgg att tta tac gcc tgg 665 Ser Ala Glu MetSer Lys Pro Ile Phe Ser Trp Ile Leu Tyr Ala Trp 120 125 130 cta cta gtgggg ctt ttt tat caa gcg cgc gtc ctg ttt gga att tta 713 Leu Leu Val GlyLeu Phe Tyr Gln Ala Arg Val Leu Phe Gly Ile Leu 135 140 145 150 ttt aagggg aaa gct caa taaatcc 738 Phe Lys Gly Lys Ala Gln 155 <210> SEQ ID NO38 <211> LENGTH: 177 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 38 Met Lys Leu Ile Lys Phe Val Arg Asn Val Val Leu PheIle Leu Thr 1 5 10 15 Ala Ile Phe Leu Ala Leu Met Leu Leu Val Ser TyrCys Met Pro His 20 25 30 Tyr Ser Val Ala Val Ile Ser Gly Val Glu Val GlnArg Met Asn Glu 35 40 45 Asn Ala Arg Pro Asn Asn Lys Glu Val Lys Thr LeuAla Arg Asp Val 50 55 60 Tyr Phe Val Gln Thr Tyr Asp Pro Lys Asp Gln LysSer Val Thr Val 65 70 75 80 Tyr Arg Asn Glu Asp Thr Arg Phe Gly Phe ProPhe Tyr Phe Lys Phe 85 90 95 Asn Ser Ala Asp Ile Ser Ala Leu Ala Gln SerLeu Val Asn Gln Gln 100 105 110 Val Glu Val Gln Tyr Tyr Gly Trp Arg IleAsn Leu Phe Asn Met Phe 115 120 125 Pro Asn Val Ile Phe Leu Lys Pro LeuLys Glu Ser Ala Glu Met Ser 130 135 140 Lys Pro Ile Phe Ser Trp Ile LeuTyr Ala Trp Leu Leu Val Gly Leu 145 150 155 160 Phe Tyr Gln Ala Arg ValLeu Phe Gly Ile Leu Phe Lys Gly Lys Ala 165 170 175 Gln <210> SEQ ID NO39 <211> LENGTH: 435 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (1)...(432) <400>SEQUENCE: 39 atg tta gaa aaa ttg att gaa aga gtg ttg ttt gcc act cgt tggttg 48 Met Leu Glu Lys Leu Ile Glu Arg Val Leu Phe Ala Thr Arg Trp Leu 15 10 15 cta gcc cct tta tgt att gcc atg tcg tta gtg ctg gtg gtt tta ggc96 Leu Ala Pro Leu Cys Ile Ala Met Ser Leu Val Leu Val Val Leu Gly 20 2530 tat gtg ttc atg aaa gag ttg tgg cac atg ctc agc cat tta aac acg 144Tyr Val Phe Met Lys Glu Leu Trp His Met Leu Ser His Leu Asn Thr 35 40 45atc agc gaa acg gat ttg gtt tta tca gcc tta gga tta gtg gat ttg 192 IleSer Glu Thr Asp Leu Val Leu Ser Ala Leu Gly Leu Val Asp Leu 50 55 60 ttgttt atg gcc ggg ctt gtt tta atg gtg tta ctc gcc agt tat gaa 240 Leu PheMet Ala Gly Leu Val Leu Met Val Leu Leu Ala Ser Tyr Glu 65 70 75 80 agcttt gtt tct aaa tta gac aag gtg gat gcc agt gaa atc act tgg 288 Ser PheVal Ser Lys Leu Asp Lys Val Asp Ala Ser Glu Ile Thr Trp 85 90 95 cta aagcac acg gat ttt aac gct tta aaa tta aag gtt tca ctc tcc 336 Leu Lys HisThr Asp Phe Asn Ala Leu Lys Leu Lys Val Ser Leu Ser 100 105 110 att gtagcg att tca gcg att ttc ttg ctc aaa cgc tac atg agt tta 384 Ile Val AlaIle Ser Ala Ile Phe Leu Leu Lys Arg Tyr Met Ser Leu 115 120 125 gaa agatgt ttt atc cca gca ttc cct aag gat acg ccc cct atc gca 432 Glu Arg CysPhe Ile Pro Ala Phe Pro Lys Asp Thr Pro Pro Ile Ala 130 135 140 taa 435<210> SEQ ID NO 40 <211> LENGTH: 144 <212> TYPE: PRT <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 40 Met Leu Glu Lys Leu Ile Glu ArgVal Leu Phe Ala Thr Arg Trp Leu 1 5 10 15 Leu Ala Pro Leu Cys Ile AlaMet Ser Leu Val Leu Val Val Leu Gly 20 25 30 Tyr Val Phe Met Lys Glu LeuTrp His Met Leu Ser His Leu Asn Thr 35 40 45 Ile Ser Glu Thr Asp Leu ValLeu Ser Ala Leu Gly Leu Val Asp Leu 50 55 60 Leu Phe Met Ala Gly Leu ValLeu Met Val Leu Leu Ala Ser Tyr Glu 65 70 75 80 Ser Phe Val Ser Lys LeuAsp Lys Val Asp Ala Ser Glu Ile Thr Trp 85 90 95 Leu Lys His Thr Asp PheAsn Ala Leu Lys Leu Lys Val Ser Leu Ser 100 105 110 Ile Val Ala Ile SerAla Ile Phe Leu Leu Lys Arg Tyr Met Ser Leu 115 120 125 Glu Arg Cys PheIle Pro Ala Phe Pro Lys Asp Thr Pro Pro Ile Ala 130 135 140 <210> SEQ IDNO 41 <211> LENGTH: 519 <212> TYPE: DNA <213> ORGANISM: Helicobacterpylori <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (1)...(516)<221> NAME/KEY: sig_peptide <222> LOCATION: (1)...(60) <221> NAME/KEY:mat_peptide <222> LOCATION: (61)...(516) <400> SEQUENCE: 41 atg cgt ttatta ttg tgg tgg gta ttg gta tta tcg ctc ttt tta aat 48 Met Arg Leu LeuLeu Trp Trp Val Leu Val Leu Ser Leu Phe Leu Asn -20 -15 -10 -5 cct ttgaga gcg gtt gaa gag cat gaa aca gat gcg gtg gat ttg ttt 96 Pro Leu ArgAla Val Glu Glu His Glu Thr Asp Ala Val Asp Leu Phe 1 5 10 ttg att ttcaat caa atc aac caa ctc aat caa gtc att gaa act tat 144 Leu Ile Phe AsnGln Ile Asn Gln Leu Asn Gln Val Ile Glu Thr Tyr 15 20 25 aag aaa aac cctgaa aga agt gct gaa atc tct ctg tat aac acc caa 192 Lys Lys Asn Pro GluArg Ser Ala Glu Ile Ser Leu Tyr Asn Thr Gln 30 35 40 aag aat gat ttg attaaa agt ttg act tct aaa gtg ttg aat gaa agg 240 Lys Asn Asp Leu Ile LysSer Leu Thr Ser Lys Val Leu Asn Glu Arg 45 50 55 60 gat aaa att ggc attgat atc aat caa aat tta aaa gag caa gag aaa 288 Asp Lys Ile Gly Ile AspIle Asn Gln Asn Leu Lys Glu Gln Glu Lys 65 70 75 atc aaa aag cgc ttg tctaga agc att aag ggc gat aat ttc tac act 336 Ile Lys Lys Arg Leu Ser ArgSer Ile Lys Gly Asp Asn Phe Tyr Thr 80 85 90 ttc atg aaa gac aga ttg tcttta gat att ttg ttg ata gat gaa att 384 Phe Met Lys Asp Arg Leu Ser LeuAsp Ile Leu Leu Ile Asp Glu Ile 95 100 105 ttg tat cgt ttt ata gat aaaatc aag agc agt att gat att ttt agc 432 Leu Tyr Arg Phe Ile Asp Lys IleLys Ser Ser Ile Asp Ile Phe Ser 110 115 120 gaa caa aaa gat gtg gaa agtttc agc gat gcc ttc ctt ttg cgt ttt 480 Glu Gln Lys Asp Val Glu Ser PheSer Asp Ala Phe Leu Leu Arg Phe 125 130 135 140 agg aca att cca act catacc ctt tcc cta aaa att taa 519 Arg Thr Ile Pro Thr His Thr Leu Ser LeuLys Ile 145 150 <210> SEQ ID NO 42 <211> LENGTH: 172 <212> TYPE: PRT<213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 42 Met Arg Leu LeuLeu Trp Trp Val Leu Val Leu Ser Leu Phe Leu Asn 1 5 10 15 Pro Leu ArgAla Val Glu Glu His Glu Thr Asp Ala Val Asp Leu Phe 20 25 30 Leu Ile PheAsn Gln Ile Asn Gln Leu Asn Gln Val Ile Glu Thr Tyr 35 40 45 Lys Lys AsnPro Glu Arg Ser Ala Glu Ile Ser Leu Tyr Asn Thr Gln 50 55 60 Lys Asn AspLeu Ile Lys Ser Leu Thr Ser Lys Val Leu Asn Glu Arg 65 70 75 80 Asp LysIle Gly Ile Asp Ile Asn Gln Asn Leu Lys Glu Gln Glu Lys 85 90 95 Ile LysLys Arg Leu Ser Arg Ser Ile Lys Gly Asp Asn Phe Tyr Thr 100 105 110 PheMet Lys Asp Arg Leu Ser Leu Asp Ile Leu Leu Ile Asp Glu Ile 115 120 125Leu Tyr Arg Phe Ile Asp Lys Ile Lys Ser Ser Ile Asp Ile Phe Ser 130 135140 Glu Gln Lys Asp Val Glu Ser Phe Ser Asp Ala Phe Leu Leu Arg Phe 145150 155 160 Arg Thr Ile Pro Thr His Thr Leu Ser Leu Lys Ile 165 170<210> SEQ ID NO 43 <211> LENGTH: 432 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION:(1)...(429) <221> NAME/KEY: sig_peptide <222> LOCATION: (1)...(93) <221>NAME/KEY: mat_peptide <222> LOCATION: (94)...(429) <400> SEQUENCE: 43atg aaa aaa ttt ttt tct caa tct tta tta gct ttg att gtg tct atg 48 MetLys Lys Phe Phe Ser Gln Ser Leu Leu Ala Leu Ile Val Ser Met -30 -25 -20aac gcg cta ctg gcc atg gat ggc aat ggc gtt ttt tta ggg gcg ggt 96 AsnAla Leu Leu Ala Met Asp Gly Asn Gly Val Phe Leu Gly Ala Gly -15 -10 -5 1tat ttg caa ggg caa gcc caa atg cat gcg gat att aat tct caa aaa 144 TyrLeu Gln Gly Gln Ala Gln Met His Ala Asp Ile Asn Ser Gln Lys 5 10 15 caagcc act aac gct act atc aaa ggc ttt gat gcg ctt tta ggg tat 192 Gln AlaThr Asn Ala Thr Ile Lys Gly Phe Asp Ala Leu Leu Gly Tyr 20 25 30 caa tttttc ttt ggg aaa tac ttt ggc ttg cgt gct tat ggg ttt ttt 240 Gln Phe PhePhe Gly Lys Tyr Phe Gly Leu Arg Ala Tyr Gly Phe Phe 35 40 45 gac tac gctcat gcc aat tct att agg ctt aaa aac cct aac tat aac 288 Asp Tyr Ala HisAla Asn Ser Ile Arg Leu Lys Asn Pro Asn Tyr Asn 50 55 60 65 agc gaa gtggcg caa ttg gcg ggt caa att ctt ggg aaa caa gaa atc 336 Ser Glu Val AlaGln Leu Ala Gly Gln Ile Leu Gly Lys Gln Glu Ile 70 75 80 aat cgc tta acgagc ctt gct gat cct aaa acc ttt gag cca aac atg 384 Asn Arg Leu Thr SerLeu Ala Asp Pro Lys Thr Phe Glu Pro Asn Met 85 90 95 ctc act tat ggg ggggct atg gat tta atg gtt aat gtt cat caa 429 Leu Thr Tyr Gly Gly Ala MetAsp Leu Met Val Asn Val His Gln 100 105 110 taa 432 <210> SEQ ID NO 44<211> LENGTH: 143 <212> TYPE: PRT <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 44 Met Lys Lys Phe Phe Ser Gln Ser Leu Leu Ala Leu IleVal Ser Met 1 5 10 15 Asn Ala Leu Leu Ala Met Asp Gly Asn Gly Val PheLeu Gly Ala Gly 20 25 30 Tyr Leu Gln Gly Gln Ala Gln Met His Ala Asp IleAsn Ser Gln Lys 35 40 45 Gln Ala Thr Asn Ala Thr Ile Lys Gly Phe Asp AlaLeu Leu Gly Tyr 50 55 60 Gln Phe Phe Phe Gly Lys Tyr Phe Gly Leu Arg AlaTyr Gly Phe Phe 65 70 75 80 Asp Tyr Ala His Ala Asn Ser Ile Arg Leu LysAsn Pro Asn Tyr Asn 85 90 95 Ser Glu Val Ala Gln Leu Ala Gly Gln Ile LeuGly Lys Gln Glu Ile 100 105 110 Asn Arg Leu Thr Ser Leu Ala Asp Pro LysThr Phe Glu Pro Asn Met 115 120 125 Leu Thr Tyr Gly Gly Ala Met Asp LeuMet Val Asn Val His Gln 130 135 140 <210> SEQ ID NO 45 <211> LENGTH: 336<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: CDS <222> LOCATION: (1)...(333) <221> NAME/KEY: sig_peptide<222> LOCATION: (1)...(60) <221> NAME/KEY: mat_peptide <222> LOCATION:(61)...(333) <400> SEQUENCE: 45 atg aaa acc ttt aaa aac ctg ctc tgt tttagc ctg atc gct atg agt 48 Met Lys Thr Phe Lys Asn Leu Leu Cys Phe SerLeu Ile Ala Met Ser -20 -15 -10 -5 tgg ctc caa gcg gac atg ttg gat aatttc act agg gcc att aac agc 96 Trp Leu Gln Ala Asp Met Leu Asp Asn PheThr Arg Ala Ile Asn Ser 1 5 10 tac acc act aaa aag ctt aat gaa atc aaggat caa gtc aat agc gct 144 Tyr Thr Thr Lys Lys Leu Asn Glu Ile Lys AspGln Val Asn Ser Ala 15 20 25 aac cct act aaa aat cac aat acc act tat aacgct aat ggc atg ctc 192 Asn Pro Thr Lys Asn His Asn Thr Thr Tyr Asn AlaAsn Gly Met Leu 30 35 40 att aac att gat tgt aaa gtc tta aaa aat aac ttctat tcg gtg tgt 240 Ile Asn Ile Asp Cys Lys Val Leu Lys Asn Asn Phe TyrSer Val Cys 45 50 55 60 tat tct agc gag tta aaa aac cct att tat ggc gtgagc gtg ttg ttt 288 Tyr Ser Ser Glu Leu Lys Asn Pro Ile Tyr Gly Val SerVal Leu Phe 65 70 75 ggg gat tta gtg gat aaa aat aat att gaa aaa cgc tatgag ttt 333 Gly Asp Leu Val Asp Lys Asn Asn Ile Glu Lys Arg Tyr Glu Phe80 85 90 taa 336 <210> SEQ ID NO 46 <211> LENGTH: 111 <212> TYPE: PRT<213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 46 Met Lys Thr PheLys Asn Leu Leu Cys Phe Ser Leu Ile Ala Met Ser 1 5 10 15 Trp Leu GlnAla Asp Met Leu Asp Asn Phe Thr Arg Ala Ile Asn Ser 20 25 30 Tyr Thr ThrLys Lys Leu Asn Glu Ile Lys Asp Gln Val Asn Ser Ala 35 40 45 Asn Pro ThrLys Asn His Asn Thr Thr Tyr Asn Ala Asn Gly Met Leu 50 55 60 Ile Asn IleAsp Cys Lys Val Leu Lys Asn Asn Phe Tyr Ser Val Cys 65 70 75 80 Tyr SerSer Glu Leu Lys Asn Pro Ile Tyr Gly Val Ser Val Leu Phe 85 90 95 Gly AspLeu Val Asp Lys Asn Asn Ile Glu Lys Arg Tyr Glu Phe 100 105 110 <210>SEQ ID NO 47 <211> LENGTH: 755 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION:(223)...(672) <221> NAME/KEY: sig_peptide <222> LOCATION: (223)...(285)<221> NAME/KEY: mat_peptide <222> LOCATION: (286)...(672) <400>SEQUENCE: 47 gattttaagc gcttgaaata gcccatttta atcaacaatt aagcgactaaccattaaact 60 taagcgataa ataagataaa atttaggata gctcaaatct ttataaaaagaaaaggataa 120 ccccttacaa actttatttt taataaaaaa tggcttatct cttctagcctactcccctta 180 ttttttctta accctttagc ggcagaagat gatggatttt tt atg ggggtg agt 234 Met Gly Val Ser -20 tat caa act tct cta gcc gtt caa agg gtggat aac tca ggg ctt aac 282 Tyr Gln Thr Ser Leu Ala Val Gln Arg Val AspAsn Ser Gly Leu Asn -15 -10 -5 gcc agt caa gac gca tcc act tac atc cgccaa aac gct atc gct cta 330 Ala Ser Gln Asp Ala Ser Thr Tyr Ile Arg GlnAsn Ala Ile Ala Leu 1 5 10 15 gaa tct gcg gca gtg cct tta gcc tat tattta gaa gcg atg ggc caa 378 Glu Ser Ala Ala Val Pro Leu Ala Tyr Tyr LeuGlu Ala Met Gly Gln 20 25 30 caa acc aga gtt tta atg caa atg ctc tgc cctgat ccg tct aaa aga 426 Gln Thr Arg Val Leu Met Gln Met Leu Cys Pro AspPro Ser Lys Arg 35 40 45 tgt ttg ctc tat gcg ggg ggt tat aaa aac gga tcaagt aat act aac 474 Cys Leu Leu Tyr Ala Gly Gly Tyr Lys Asn Gly Ser SerAsn Thr Asn 50 55 60 ggc gat aca ggc aac aac ccc cca aga ggc aat gtc aatgcc acc ttt 522 Gly Asp Thr Gly Asn Asn Pro Pro Arg Gly Asn Val Asn AlaThr Phe 65 70 75 gat atg caa tct tta gtc aat aat cta aac aaa ctc acc caactc atc 570 Asp Met Gln Ser Leu Val Asn Asn Leu Asn Lys Leu Thr Gln LeuIle 80 85 90 95 ggc gag act tta atc cgt aac cct gaa aat ctt tct aac gccaaa gtc 618 Gly Glu Thr Leu Ile Arg Asn Pro Glu Asn Leu Ser Asn Ala LysVal 100 105 110 ttt aac gtc aaa ttt gga aat caa agc act gtt att gct tttacc aga 666 Phe Asn Val Lys Phe Gly Asn Gln Ser Thr Val Ile Ala Phe ThrArg 115 120 125 ggg tct tagacaaata ccatgggacg cttttacaaa tgacaatcaaccaacgcttt 722 Gly Ser taaccacgct ctgggtattt accaaaaccc taa 755 <210>SEQ ID NO 48 <211> LENGTH: 150 <212> TYPE: PRT <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 48 Met Gly Val Ser Tyr Gln Thr SerLeu Ala Val Gln Arg Val Asp Asn 1 5 10 15 Ser Gly Leu Asn Ala Ser GlnAsp Ala Ser Thr Tyr Ile Arg Gln Asn 20 25 30 Ala Ile Ala Leu Glu Ser AlaAla Val Pro Leu Ala Tyr Tyr Leu Glu 35 40 45 Ala Met Gly Gln Gln Thr ArgVal Leu Met Gln Met Leu Cys Pro Asp 50 55 60 Pro Ser Lys Arg Cys Leu LeuTyr Ala Gly Gly Tyr Lys Asn Gly Ser 65 70 75 80 Ser Asn Thr Asn Gly AspThr Gly Asn Asn Pro Pro Arg Gly Asn Val 85 90 95 Asn Ala Thr Phe Asp MetGln Ser Leu Val Asn Asn Leu Asn Lys Leu 100 105 110 Thr Gln Leu Ile GlyGlu Thr Leu Ile Arg Asn Pro Glu Asn Leu Ser 115 120 125 Asn Ala Lys ValPhe Asn Val Lys Phe Gly Asn Gln Ser Thr Val Ile 130 135 140 Ala Phe ThrArg Gly Ser 145 150 <210> SEQ ID NO 49 <211> LENGTH: 29 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 49 gccngaagga tttattatga ttaaaagaa 29 <210> SEQ ID NO 50<211> LENGTH: 29 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 50 gccnaaggaataaattagaa agtgaagaa 29 <210> SEQ ID NO 51 <211> LENGTH: 25 <212> TYPE:DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 51 gccnaaaggg cgaaaatgag caaga 25 <210> SEQ ID NO 52<211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 52 gccnttttttaagaatcact ttcttcgg 28 <210> SEQ ID NO 53 <211> LENGTH: 27 <212> TYPE:DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 53 gccncgcatt gatttgatga ataaacc 27 <210> SEQ ID NO 54<211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 54 gccnctcactaaaaagcaat ttttgag 27 <210> SEQ ID NO 55 <211> LENGTH: 28 <212> TYPE:DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 55 gccntcacaa tggataaaaa caacaaca 28 <210> SEQ ID NO 56<211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 56 gccnctgtccaaatcagcca ccc 23 <210> SEQ ID NO 57 <211> LENGTH: 25 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 57 gccnggaaga ataatgctcg cttcc 25 <210> SEQ ID NO 58<211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 58 gccnctattctccagggata tggcc 25 <210> SEQ ID NO 59 <211> LENGTH: 28 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 59 gccngaaggg tgtatggtat taggaagc 28 <210> SEQ ID NO 60<211> LENGTH: 29 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 60 gccncgttaaaactaaagtt ctattttta 29 <210> SEQ ID NO 61 <211> LENGTH: 28 <212> TYPE:DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 61 gccnaatata tgggaactta atgagaat 28 <210> SEQ ID NO 62<211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 62 gccnatgtcatgtcaaacta tgaagc 26 <210> SEQ ID NO 63 <211> LENGTH: 27 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 63 gccnaaaagg gttttaaata atggctg 27 <210> SEQ ID NO 64<211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 64 gccnaagattctaaaagggc ttcaaat 27 <210> SEQ ID NO 65 <211> LENGTH: 38 <212> TYPE:DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 65 gccngagagt agtggcagag tttatgctga ttccgtta 38 <210>SEQ ID NO 66 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 66gccnggctta aactggaacg gatttc 26 <210> SEQ ID NO 67 <211> LENGTH: 30<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 67 gccnatgaaa agatttgatt tgtttttatc 30 <210>SEQ ID NO 68 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 68gccnttaaat atcccaatcc tgccac 26 <210> SEQ ID NO 69 <211> LENGTH: 35<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 69 gccntaaagt ttgctaaaaa gatggtttta atttc 35<210> SEQ ID NO 70 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 70gccncccatc tttagaaatc aaccccca 28 <210> SEQ ID NO 71 <211> LENGTH: 32<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 71 gccncaataa aacaccaaaa tgaatgagtt ac 32<210> SEQ ID NO 72 <211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 72gccngcattt accccctaaa aactataaac 30 <210> SEQ ID NO 73 <211> LENGTH: 31<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 73 gccngtaagg aatgagatga taaagagttg g 31<210> SEQ ID NO 74 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 74gccnctaaac tctggcttat tgcgtatc 28 <210> SEQ ID NO 75 <211> LENGTH: 30<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 75 gccnatagga acaagcatgt tttttaaaac 30 <210>SEQ ID NO 76 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 76gccnctggct tattgcgtat catc 24 <210> SEQ ID NO 77 <211> LENGTH: 32 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 77 gccngaaatc aaggagtttg tatgcaacag cg 32<210> SEQ ID NO 78 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 78gccncccaat acttttattg attcaccatt tc 32 <210> SEQ ID NO 79 <211> LENGTH:27 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE:<221> NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION:n = A,T,C or G <400> SEQUENCE: 79 gccnatggta tttgacagaa caatcag 27 <210>SEQ ID NO 80 <211> LENGTH: 29 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 80gccnttagga atagcataac aaacaaacg 29 <210> SEQ ID NO 81 <211> LENGTH: 29<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 81 gccnatgttg aaatttaaat atggtttga 29 <210>SEQ ID NO 82 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 82gccngagcct acaggttgct tgc 23 <210> SEQ ID NO 83 <211> LENGTH: 31 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 83 gccncaagca aaaaaatgtt caataaaagg g 31<210> SEQ ID NO 84 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 84gccngtctaa attagaataa gtgttgtt 28 <210> SEQ ID NO 85 <211> LENGTH: 30<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 85 gccntaagga atgaagttga taaaatttgt 30 <210>SEQ ID NO 86 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 86gccnggattt attgagcttt cccctt 26 <210> SEQ ID NO 87 <211> LENGTH: 29<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 87 gccnatgtta gaaaaattga ttgaaagag 29 <210>SEQ ID NO 88 <211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 88gccnttatgc gatagggggc gtatc 25 <210> SEQ ID NO 89 <211> LENGTH: 26 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221>NAME/KEY: misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n =A,T,C or G <400> SEQUENCE: 89 gccnatgcgt ttattattgt ggtggg 26 <210> SEQID NO 90 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Helicobacterpylori <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4<223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 90 gccnttaaatttttagggaa agggta 26 <210> SEQ ID NO 91 <211> LENGTH: 30 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 91 gccnatgaaa aaattttttt ctcaatcttt 30 <210> SEQ ID NO92 <211> LENGTH: 29 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 92 gccnttattgatgaacatta accattaaa 29 <210> SEQ ID NO 93 <211> LENGTH: 26 <212> TYPE:DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 93 gccnatgaaa acctttaaaa acctgc 26 <210> SEQ ID NO 94<211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 94 gccnttaaaactcatagcgt ttttcaat 28 <210> SEQ ID NO 95 <211> LENGTH: 27 <212> TYPE:DNA <213> ORGANISM: Helicobacter pylori <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: 4 <223> OTHER INFORMATION: n = A,T,C or G<400> SEQUENCE: 95 gccngatgga ttttttatgg gggtgag 27 <210> SEQ ID NO 96<211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 4 <223>OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 96 gccnatggtatttgtctaag accctc 26 <210> SEQ ID NO 97 <211> LENGTH: 24 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 97 aacctaatttgaaattcaaa ccat 24 <210> SEQ ID NO 98 <211> LENGTH: 32 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 98 aaattaggttttgtaggctt tgccaataaa tg 32 <210> SEQ ID NO 99 <211> LENGTH: 23 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 99taaaataacc aacagagtga tca 23 <210> SEQ ID NO 100 <211> LENGTH: 33 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 100ggttatttta gtggatattt gggtttatag cga 33 <210> SEQ ID NO 101 <211>LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 101 cgcctataac cgctccatt 19 <210> SEQ ID NO 102 <211> LENGTH:32 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE:102 gttataggcg ataaaggttt aacgcagcta ag 32 <210> SEQ ID NO 103 <211>LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 103 gcccttttgt ttaggggtta g 21 <210> SEQ ID NO 104 <211>LENGTH: 32 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 104 acaaaagggc tttttagagc atgtgagcca tc 32 <210> SEQ ID NO 105<211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 105 actggagtgt ggataaaact at 22 <210> SEQ ID NO 106<211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 106 acactccagt agatgctttc ccggatattt c 31 <210> SEQ IDNO 107 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Helicobacterpylori <400> SEQUENCE: 107 caccatacat gtatcctgca ttaatg 26 <210> SEQ IDNO 108 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Helicobacterpylori <400> SEQUENCE: 108 catgtatggt gtagcaaaga attttaagga ggc 33 <210>SEQ ID NO 109 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 109 tgcgagattt aacctgtttt ca 22<210> SEQ ID NO 110 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 110 aaatctcgca gaaatctttc acaagcgagcaa 32 <210> SEQ ID NO 111 <211> LENGTH: 22 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 111 acaaggataa aaaacgcgctaa 22 <210> SEQ ID NO 112 <211> LENGTH: 33 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 112 ttatccttgt tgctggcttggtttttttta att 33 <210> SEQ ID NO 113 <211> LENGTH: 30 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 113 aacttttctctatcccaatt cgttacgctc 30 <210> SEQ ID NO 114 <211> LENGTH: 31 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 114ggatagagaa aagtttggcg tcaaaagttg g 31 <210> SEQ ID NO 115 <211> LENGTH:22 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE:115 aagccgtatt gtttgttttg gc 22 <210> SEQ ID NO 116 <211> LENGTH: 34<212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 116aatacggctt taaagctata gaaaatttaa acgc 34 <210> SEQ ID NO 117 <211>LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 117 gacttctaaa gcgtcctttt tttcttta 28 <210> SEQ ID NO 118<211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 118 ctttagaagt cattaaacaa agaggggt 28 <210> SEQ ID NO119 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 119 agattttgtt ttgagcgtta gaaatg 26 <210> SEQ ID NO 120<211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 120 caaaatctat aaactcaatc aagtcaaaaa tg 32 <210> SEQ IDNO 121 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Helicobacterpylori <400> SEQUENCE: 121 tggaatattg tgatccacgc catc 24 <210> SEQ ID NO122 <211> LENGTH: 37 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 122 gaatattcca aaagccgttt tttattacag aagaggg 37 <210>SEQ ID NO 123 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 123 tgaagtcttg cgatttttgc tt 22<210> SEQ ID NO 124 <211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 124 caagacttca aaaaagaagg agcggttgcc30 <210> SEQ ID NO 125 <211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 125 aagcttttca ttatcttccc cataagc 27<210> SEQ ID NO 126 <211> LENGTH: 29 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 126 tgaaaagctt ttagcgaagc gatcaagcc29 <210> SEQ ID NO 127 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 127 gaaaagccac cccgcttatt 20 <210>SEQ ID NO 128 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM:Helicobacter pylori <400> SEQUENCE: 128 gtggcttttc aaaaagagtg ggtgcaacaatt 32 <210> SEQ ID NO 129 <211> LENGTH: 21 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 129 aaaccccact cttatcatcgg 21 <210> SEQ ID NO 130 <211> LENGTH: 30 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 130 agtggggttt ttttagggggtgggtatgct 30 <210> SEQ ID NO 131 <211> LENGTH: 22 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 131 taagtccata cgatagcctatg 22 <210> SEQ ID NO 132 <211> LENGTH: 34 <212> TYPE: DNA <213>ORGANISM: Helicobacter pylori <400> SEQUENCE: 132 tatggaactt agaacattttaacacgctct atta 34 <210> SEQ ID NO 133 <211> LENGTH: 23 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 133 gcattttcattcattctttg gac 23 <210> SEQ ID NO 134 <211> LENGTH: 32 <212> TYPE: DNA<213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 134 atgaaaatgcacgcccaaat aataaggaag ta 32 <210> SEQ ID NO 135 <211> LENGTH: 22 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 135tgaacacata gcctaaaacc ac 22 <210> SEQ ID NO 136 <211> LENGTH: 31 <212>TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE: 136tatgtgttca tgaaagagtt gtggcacatg c 31 <210> SEQ ID NO 137 <211> LENGTH:27 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE:137 caatacccac cacaataata aacgcat 27 <210> SEQ ID NO 138 <211> LENGTH:33 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE:138 gtgggtattg gtattatcgc tctttttaaa tcc 33 <210> SEQ ID NO 139 <211>LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 139 tggccagtag cgcgttcat 19 <210> SEQ ID NO 140 <211> LENGTH:34 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400> SEQUENCE:140 ctactggcca tggatggcaa tggcgttttt ttag 34 <210> SEQ ID NO 141 <211>LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 141 tagcgatcag gctaaaacag a 21 <210> SEQ ID NO 142 <211>LENGTH: 29 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 142 ctgatcgcta tgagttggct ccaagcgga 29 <210> SEQ ID NO 143<211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 143 ggcactgccg cagattcta 19 <210> SEQ ID NO 144 <211>LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori <400>SEQUENCE: 144 cggcagtgcc tttagcctat tatttagaag cga 33 <210> SEQ ID NO145 <211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 145 gccgagctct atcgtatgga cttagaacat 30 <210> SEQ ID NO146 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Helicobacter pylori<400> SEQUENCE: 146 gccctcgaga ttagaataag tgttgtttaa aatc 34

What is claimed is:
 1. An isolated polynucleotide that encodes (i) apolypeptide comprising an amino acid sequence that is homologous to theamino acid sequence of a Helicobacter membrane-associated polypeptide,wherein said amino acid sequence of said Helicobactermembrane-associated polypeptide is selected from the group consistingof: (a) the amino acid sequences as shown: in SEQ ID NO:2, beginningwith an amino acid in any one of the positions from −27 to 5, and endingwith an amino acid in position 160 (HPO101); in SEQ ID NO:4, beginningwith an amino acid in position 1 and ending with an amino acid inposition 172 (HPO104); in SEQ ID NO:6, beginning with an amino acid inany one of the positions from −17 to 5, and ending with an amino acid inposition 169 (HPO116); in SEQ ID NO:8, beginning with an amino acid inany one of the positions from −21 to 5, and ending with an amino acid inposition 198 (HPO121); in SEQ ID NO:10, beginning with an amino acid inany one of the positions from −20 to 5, and ending with an amino acid inposition 132 (HPO132); in SEQ ID NO:12, beginning with an amino acid inpositions 1 and ending with an amino acid in position 114 (HPO15); inSEQ ID NO:14, beginning with an amino acid in any one of the positionsfrom −17 to 5, and ending with an amino acid in position 248 (HPO18); inSEQ ID NO:16, beginning with an amino acid in any one of the positionsfrom −40 to 5, and ending with an amino acid in position 74 (HPO38); inSEQ ID NO:18, beginning with an amino acid in any one of the positionsfrom −34 to 5, and ending with an amino acid in position 226 (HPO42); inSEQ ID NO:20, beginning with an amino acid in any one of the positionsfrom −21 to 5, and ending with an amino acid in position 179 (HPO45); inSEQ ID NO:22, beginning with an amino acid in any one of the positionsfrom −33 to 5, and ending with an amino acid in position 114 (HPO50); inSEQ ID NO:24, beginning with an amino acid in any one of the positionsfrom −60 to 5, and ending with an amino acid in position 349 (HPO54); inSEQ ID NO:26, beginning with an amino acid in any one of the positionsfrom −18 to 5, and ending with an amino acid in position 288 (HPO57); inSEQ ID NO:28, beginning with an amino acid in any one of the positionsfrom −21 to 5, and ending with an amino acid in position 150 (HPO58); inSEQ ID NO:30, beginning with an amino acid in any one of the positionsfrom −20 to 5, and ending with an amino acid in position 309 (HPO64); inSEQ ID NO:32, beginning with an amino acid in any one of the positionsfrom −35 to 5, and ending with an amino acid in position 129 (HPO70); inSEQ ID NO:34, beginning with an amino acid in any one of the positionsfrom −19 to 5, and ending with an amino acid in position 153 (HPO71); inSEQ ID NO:36, beginning with an amino acid in any one of the positionsfrom −25 to 5, and ending with an amino acid in position 176 (HPO76); inSEQ ID NO:38, beginning with an amino acid in any one of the positionsfrom −21 to 5, and ending with an amino acid in position 156 (HPO7); inSEQ ID NO:40, beginning with an amino acid in position 1 and ending withan amino acid in position 144 (HPO80); in SEQ ID NO:42, beginning withan amino acid in any one of the positions from −20 to 5, and ending withan amino acid in position 152 (HPO87); in SEQ ID NO:44, beginning withan amino acid in any one of the positions from −31 to 5, and ending withan amino acid in position 112 (HPO95); in SEQ ID NO:46, beginning withan amino acid in any one of the positions from −20 to 5, and ending withan amino acid in position 91 (HPO98); in SEQ ID NO:48, beginning with anamino acid in any one of the positions from −21 to 5, and ending with anamino acid in position 129 (HPO9); and (b) the precursor or mature aminoacid sequences encoded by the Helicobacter DNA inserts found in AmericanType Culture Collection deposit numbers 98197 (HPO76), 98210 (HPO18),98201 (HPO121), 98208 (HPO45), 98198 (HPO101), 98200 (HPO116), 98211(HPO7), 98199 (HPO104), 98214 (HPO15), 98206 (HPO58), 98202 (HPO132),98203 (HPO9), 98204 (HPO38), 98205 (HPO87), 98217 (HPO71), 98219(HPO70), 98215 (HPO80), 98216 (HPO95), 98218 (HPO98), 98220 (HPO57),98207 (HPO50), 98213 (HPO64), 98212 (HPO54), and 98209 (HPO42); or (ii)a derivative of said polypeptide encoded by said polynucleotide.
 2. Anisolated polynucleotide that encodes (i) a polypeptide comprising anamino acid sequence that is homologous to an amino acid sequenceselected from the group consisting of: (a) the amino acid sequences asshown: in SEQ ID NO:2, beginning with an amino acid in position −27 andending with an amino acid in position 160 (HPO101); in SEQ ID NO:4,beginning with an amino acid in position 1 and ending with an amino acidin position 172 (HPO104); in SEQ ID NO:6, beginning with an amino acidin position −17 and ending with an amino acid in position 169 (HPO116);in SEQ ID NO:8, beginning with an amino acid in position −21 and endingwith an amino acid in position 198 (HPO121); in SEQ ID NO:10, beginningwith an amino acid in position −20, and ending with an amino acid inposition 132 (HPO132); in SEQ ID NO:12, beginning with an amino acid inposition 1 and ending with an amino acid in position 114 (HPO15); in SEQID NO:14, beginning with an amino acid in position −17 and ending withan amino acid in position 248 (HPO18); in SEQ ID NO:16, beginning withan amino acid in position −40 and ending with an amino acid in position74 (HPO38); in SEQ ID NO:18, beginning with an amino acid in position−34 and ending with an amino acid in position 226 (HPO42); in SEQ IDNO:20, beginning with an amino acid in position −21 and ending with anamino acid in position 179 (HPO45); in SEQ ID NO:22, beginning with anamino acid in position −33 and ending with an amino acid in position 114(HPO50); in SEQ ID NO:24, beginning with an amino acid in position −60and ending with an amino acid in position 349 (HPO54); in SEQ ID NO:26,beginning with an amino acid in position −18 and ending with an aminoacid in position 288 (HPO57); in SEQ ID NO:28, beginning with an aminoacid in position −21 and ending with an amino acid in position 150(HPO58); in SEQ ID NO:30, beginning with an amino acid in position −20and ending with an amino acid in position 309 (HPO64); in SEQ ID NO:32,beginning with an amino acid in position −35 and ending with an aminoacid in position 129 (HPO70); in SEQ ID NO:34, beginning with an aminoacid in position −19 and ending with an amino acid in position 153(HPO71); in SEQ ID NO:36, beginning with an amino acid in position −25and ending with an amino acid in position 176 (HPO76); in SEQ ID NO:38,beginning with an amino acid in position −21 and ending with an aminoacid in position 156 (HPO7); in SEQ ID NO:40, beginning with an aminoacid in position 1 and ending with an amino acid in position 144(HPO80); in SEQ ID NO:42, beginning with an amino acid in position −20and ending with an amino acid in position 152 (HPO87); in SEQ ID NO:44,beginning with an amino acid in position −31 and ending with an aminoacid in position 112 (HPO95); in SEQ ID NO:46, beginning with an aminoacid in position −20 and ending with an amino acid in position 91(HPO98); in SEQ ID NO:48, beginning with an amino acid in position −21and ending with an amino acid in position 129 (HPO9); and (b) the aminoacid sequences encoded by the DNA inserts found in American Type CultureCollection deposit numbers 98197 (HPO76), 98210 (HPO18), 98201 (HPO121),98208 (HPO45), 98198 (HPO101), 98200 (HPO116), 98211 (HPO7), 98199(HPO104), 98214 (HPO15), 98206 (HPO58), 98202 (HPO132), 98203 (HPO9),98204 (HPO38), 98205 (HPO87), 98217 (HPO71), 98219 (HPO70), 98215(HPO80), 98216 (HPO95), 98218 (HPO98), 98220 (HPO57), 98207 (HPO50),98213 (HPO64), 98212 (HPO54), and 98209 (HPO42); or (ii) a derivative ofsaid polypeptide.
 3. The isolated polynucleotide of claim 1, whichencodes the mature form of (i) a polypeptide comprising an amino acidsequence that is homologous to an amino acid sequence selected from thegroup consisting of: (a) the amino acid sequences as shown: in SEQ IDNO:2, beginning with an amino acid in position −27 and ending with anamino acid in position 160 (HPO101); in SEQ ID NO:4, beginning with anamino acid in position 1 and ending with an amino acid in position 172(HPO104); in SEQ ID NO:6, beginning with an amino acid in position −17and ending with an amino acid in position 169 (HPO116); in SEQ ID NO:8,beginning with an amino acid in position −21 and ending with an aminoacid in position 198 (HPO121); in SEQ ID NO:10, beginning with an aminoacid in position −20, and ending with an amino acid in position 132(HPO132); in SEQ ID NO:12, beginning with an amino acid in position 1and ending with an amino acid in position 114 (HPO15); in SEQ ID NO:14,beginning with an amino acid in position −17 and ending with an aminoacid in position 248 (HPO18); in SEQ ID NO:16, beginning with an aminoacid in position −40 and ending with an amino acid in position 74(HPO38); in SEQ ID NO:18, beginning with an amino acid in position −34and ending with an amino acid in position 229 (HPO42); in SEQ ID NO:20,beginning with an amino acid in position −21 and ending with an aminoacid in position 179 (HPO45); in SEQ ID NO:22, beginning with an aminoacid in position −33 and ending with an amino acid in position 114(HPO50); in SEQ ID NO:24, beginning with an amino acid in position −60and ending with an amino acid in position 349 (HPO54); in SEQ ID NO:26,beginning with an amino acid in position −18 and ending with an aminoacid in position 288 (HPO57); in SEQ ID NO:28, beginning with an aminoacid in position −21 and ending with an amino acid in position 150(HPO58); in SEQ ID NO:30, beginning with an amino acid in position −20and ending with an amino acid in position 309 (HPO64); in SEQ ID NO:32,beginning with an amino acid in position −35 and ending with an aminoacid in position 129 (HPO70); in SEQ ID NO:34, beginning with an aminoacid in position −19 and ending with an amino acid in position 153(HPO71); in SEQ ID NO:36, beginning with an amino acid in position −25and ending with an amino acid in position 176 (HPO76); in SEQ ID NO:38,beginning with an amino acid in position −21 and ending with an aminoacid in position 156 (HPO7); in SEQ ID NO:40, beginning with an aminoacid in position 1 and ending with an amino acid in position 144 (HPO80); in SEQ ID NO:42, beginning with an amino acid in position −20 andending with an amino acid in position 152 (HPO 87); in SEQ ID NO:44,beginning with an amino acid in position −31 and ending with an aminoacid in position 112 (HPO 95); in SEQ ID NO:46, beginning with an aminoacid in position −20 and ending with an amino acid in position 91 (HPO98); in SEQ ID NO:48, beginning with an amino acid in position −21 andending with an amino acid in position 129 (HPO 9); and (b) the aminoacid sequences encoded by the DNA inserts found in American Type CultureCollection deposit numbers 98197 (HPO76), 98210 (HPO18), 98201 (HPO121),98208 (HPO45), 98198 (HPO101), 98200 (HPO116), 98211 (HPO7), 98199(HPO104), 98214 (HPO15), 98206 (HPO58), 98202 (HPO132), 98203 (HPO9),98204 (HPO38), 98205 (HPO87), 98217 (HPO71), 98219 (HPO70), 98215(HPO80), 98216 (HPO95), 98218 (HPO98), 98220 (HPO57), 98207 (HPO50),98213 (HPO64), 98212 (HPO54), and 98209 (HPO42); or (ii) a derivative ofsaid polypeptide.
 4. The isolated polynucleotide of claim 1, wherein thepolynucleotide is a DNA molecule.
 5. The isolated polynucleotide ofclaim 1, which is a DNA molecule that can be amplified and/or cloned bypolymerase chain reaction from an Helicobacter genome, using either: A5′ oligonucleotide primer having a sequence as shown in SEQ ID NO:49wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:50 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:51wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:52 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:53wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:54 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:55wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:56 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:57wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:58 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:59wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:60 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:61wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:62 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:63wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:64 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:65wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:66 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:67wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:68 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:69wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:70 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:71wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:72 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:73wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:74 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:75wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:76 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:77wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:78 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:79wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:80 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:81wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:82 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:83wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:84 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:85wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:86 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:87wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:88 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:89wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:90 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:91wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:93 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:95wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:94 wherein N is a restriction site; A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:97wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:96 wherein N is a restriction site; or A 5′oligonucleotide primer having a sequence as shown in SEQ ID NO:99wherein N is a restriction site, and a 3′ oligonucleotide primer havinga sequence in SEQ ID NO:98 wherein N is a restriction site.
 6. Theisolated DNA molecule of claim 5, which can be amplified and/or clonedby the polymerase chain reaction from a Helicobacter pylori genome. 7.The isolated polynucleotide of claim 1, which is a DNA molecule thatencodes the mature form or a derivative of a polypeptide encoded by theDNA molecule of claim
 5. 8. The isolated polynucleotide of claim 1,which is a DNA molecule that encodes the mature form or a derivative ofa polypeptide encoded by the DNA molecule of claim
 6. 9. A compound, ina substantially purified form, that is the mature form or a derivativeof a polypeptide comprising an amino acid sequence that is homologous toan amino acid sequence of a polypeptide associated with the Helicobactermembrane, which is selected from the group consisting of: (a) the aminoacid sequences as shown: in SEQ ID NO:2, beginning with an amino acid inposition −27 and ending with an amino acid in position 160 (HPO101); inSEQ ID NO:4, beginning with an amino acid in position 1 and ending withan amino acid in position 172 (HPO104); in SEQ ID NO:6, beginning withan amino acid in position −17 and ending with an amino acid in position169 (HPO116); in SEQ ID NO:8, beginning with an amino acid in position−21 and ending with an amino acid in position 198 (HPO121); in SEQ IDNO:10, beginning with an amino acid in position −20, and ending with anamino acid in position 132 (HPO132); in SEQ ID NO:12, beginning with anamino acid in position 1 and ending with an amino acid in position 114(HPO15); in SEQ ID NO:14, beginning with an amino acid in position −17and ending with an amino acid in position 248 (HPO18); in SEQ ID NO:16,beginning with an amino acid in position −40 and ending with an aminoacid in position 74 (HPO38); in SEQ ID NO:18, beginning with an aminoacid in position −31 and ending with an amino acid in position 226(HPO42); in SEQ ID NO:20, beginning with an amino acid in position −21and ending with an amino acid in position 179 (HPO45); in SEQ ID NO:22,beginning with an amino acid in position −33 and ending with an aminoacid in position 114 (HPO50); in SEQ ID NO:24, beginning with an aminoacid in position −60 and ending with an amino acid in position 349(HPO54); in SEQ ID NO:26, beginning with an amino acid in position −18and ending with an amino acid in position 288 (HPO57); in SEQ ID NO:28,beginning with an amino acid in position −21 and ending with an aminoacid in position 150 (HPO58); in SEQ ID NO:30, beginning with an aminoacid in position −20 and ending with an amino acid in position 309(HPO64); in SEQ ID NO:32, beginning with an amino acid in position −35and ending with an amino acid in position 129 (HPO70); in SEQ ID NO:34,beginning with an amino acid in position −19 and ending with an aminoacid in position 153 (HPO71); in SEQ ID NO:36, beginning with an aminoacid in position −25 and ending with an amino acid in position 176(HPO76); in SEQ ID NO:38, beginning with an amino acid in position −21and ending with an amino acid in position 156 (HPO7); in SEQ ID NO:40,beginning with an amino acid in position 1 and ending with an amino acidin position 144 (HPO80); in SEQ ID NO:42, beginning with an amino acidin position −20 and ending with an amino acid in position 152 (HPO87);in SEQ ID NO:44, beginning with an amino acid in position −31 and endingwith an amino acid in position 112 (HPO95); in SEQ ID NO:46, beginningwith an amino acid in position −20 and ending with an amino acid inposition 91 (HPO98); in SEQ ID NO:48, beginning with an amino acid inposition −21 and ending with an amino acid in position 129 (HPO9); and(b) the amino acid sequences encoded by the Helicobacter DNA insertsfound in American Type Culture Collection deposit numbers 98197 (HPO76),98210 (HPO18), 98201 (HPO121), 98208 (HPO45), 98198 (HPO101), 98200(HPO116), 98211 (HPO7), 98199 (HPO104), 98214 (HPO15), 98206 (HPO58),98202 (HPO132), 98203 (HPO9), 98204 (HPO38), 98205 (HPO87), 98217(HPO71), 98219 (HPO70), 98215 (HPO80), 98216 (HPO95), 98218 (HPO98),98220 (HPO57), 98207 (HPO50), 98213 (HPO64), 98212 (HPO54), and 98209(HPO42).
 10. The compound of claim 9, which is the mature form or aderivative of a polypeptide encoded by a DNA molecule of claim
 5. 11.The compound of claim 9, which is the mature form or a derivative of apolypeptide encoded by a DNA molecule of claim
 6. 12. A method ofpreventing or treating Helicobacter infection in a mammal, said methodcomprising administering to said mammal a prophylactically ortherapeutically effective amount of a compound of claim
 9. 13. Themethod of claim 12, further comprising administering an antibiotic, anantisecretory agent, a bismuth salt, or a combination thereof.
 14. Themethod of claim 13, wherein said antibiotic is selected from the groupconsisting of amoxicillin, clarithromycin, tetracycline, metronidizole,and erythromycin.
 15. The method of claim 13, wherein said bismuth saltis selected from the group consisting of bismuth subcitrate and bismuthsubsalicylate.
 16. The method of claim 13, wherein said antisecretoryagent is a proton pump inhibitor.
 17. The method of claim 16, whereinsaid proton pump inhibitor is selected from the group consisting ofomeprazole, lansoprazole, and pantoprazole.
 18. The method of claim 13,wherein said antisecretory agent is an H₂-receptor antagonist.
 19. Themethod of claim 18, wherein said H₂-receptor antagonist is selected fromthe group consisting of ranitidine, cimetidine, famotidine, nizatidine,and roxatidine.
 20. The method of claim 13, wherein said antisecretoryagent is a prostaglandin analog.
 21. The method of claim 20, whereinsaid prostaglandin analog is misoprostil or enprostil.
 22. The method ofclaim 12, which further comprises administering a prophylactically ortherapeutically effective amount of a second Helicobacter polypeptide ora derivative thereof.
 23. The method of claim 22, wherein the secondHelicobacter polypeptide is a Helicobacter urease, a subunit, or aderivative thereof.
 24. A composition comprising a compound of claim 9,together with a physiologically acceptable diluent or carrier.
 25. Thecomposition of claim 24, further comprising an adjuvant.
 26. Thecomposition of claim 24, further comprising a second Helicobacterpolypeptide or a derivative thereof.
 27. The composition of claim 26,wherein said second Helicobacter polypeptide is a Helicobacter urease,or a subunit or a derivative thereof.
 28. A method of preventing ortreating Helicobacter infection in a mammal, said method comprisingadministering to said mammal a prophylactically or therapeuticallyeffective amount of a polynucleotide of claim
 1. 29. A method ofpreventing or treating Helicobacter infection in a mammal, said methodcomprising administering to said mammal a prophylactically ortherapeutically effective amount of a polynucleotide of claim
 5. 30. Amethod of preventing or treating Helicobacter infection in a mammal,said method comprising administering to said mammal a prophylacticallyor therapeutically effective amount of a polynucleotide of claim
 8. 31.A composition comprising a viral vector, in the genome of which isinserted a DNA molecule of claim 4, said DNA molecule being placed underconditions for expression in a mammalian cell and said viral vectorbeing admixed with a physiologically acceptable diluent or carrier. 32.The composition of claim 31, wherein said viral vector is a pox virus.33. A composition that comprises a bacterial vector comprising a DNAmolecule of claim 4, said DNA molecule being placed under conditions forexpression and said bacterial vector being admixed with aphysiologically acceptable diluent or carrier.
 34. The composition ofclaim 33, wherein said vector is selected from the group consisting ofShigella, Salmonella, Vibrio cholerae, Lactobacillus, Bacille bilié deCalmette-Guérin, and Streptococcus.
 35. A composition comprising apolynucleotide of claim 1, together with a physiologically acceptablediluent or carrier.
 36. The composition of claim 35, wherein saidpolynucleotide is a DNA molecule that is inserted in a plasmid that isunable to replicate and to substantially integrate in a mammalian genomeand is placed under conditions for expression in a mammalian cell. 37.An expression cassette comprising a DNA molecule of claim 4, said DNAmolecule being placed under conditions for expression in a procaryoticor eucaryotic cell.
 38. A process for producing a compound of claim 9,which comprises culturing a procaryotic or eucaryotic cell transformedor transfected with an expression cassette of claim 37, and recoveringsaid compound from the cell culture.
 39. A method of preventing ortreating Helicobacter infection in a mammal, said method comprisingadministering to said mammal a prophylactically or therapeuticallyeffective amount of an antibody that binds to the compound of claim 9.